#Synth DNA Modification
Text
Thinking about the spectrum of what cyborgs can be for a setting I'm working on. Noodling around with a bunch of related concepts.
Prosthetic Wearer - human with artificial accessories to replicate a more 'standard' body. sometimes for utility reasons(i,e; metal arm to grab stuff), sometimes for social ones(i,e; fake nose to stop ppl staring).
Assistive Device User - human with artificial accessories that are so integral to their life that they basically count them as part of their own body, but are NOT designed to mimic a normative body plan. (wheelchair users, glasses users, etc)
Body Modder - human who chooses to deviate from a normative body. includes both organic modification(hormone therapy, scarification) and inorganic(piercings, implanted chips)
Clone type 1(realistic) - human with DNA identical to someone else(see also: identical twin) (Dolly the Sheep)
Clone type 2(cartoon) - copy of someone grown in a big vat. usually "not a real person" whatever that means. (Commander Cody)
Cyborg Type 1(fancy prosthetics) - a human with body parts that have extra gizmos built in. sometimes started as disabled, but sometimes chose to replace standard parts to get the cool gizmos (Darth Vader)
Cyborg Type 2(a new cybernetic being) - a person who has human parts, and has robot parts, and isn't really either one. May have memories of one or more human lives, and also memories and traits like a computer, but isn't just 'a person with metal bits' OR 'a robot with flesh bits'(Motoko Kusanagi)
Cyborg Type 3(meaty robot) - a robot that also has some organic parts, but was never an unmodified human. may or may not be "a real person" depending on the setting. (The Terminator)
Bioroid - a robot that's assembled from 100% organic parts. usually looks like a human, but sometimes they're animals or monsters. (gen 3 synths from Fallout)
Android Type 1(indistinguishable from human) - a robot that's built to look like a human. no organic parts, but is designed to appear organic at first glance. (2B, Rick Deckard)
Android Type 2(loosely human shaped robot) - A robot that looks kind of like a human, but is obvious artificial. (C3po, V1)
Transformer - a robot that only looks human sometimes. (Optimus Prime)
Robot - a machine that passes some threshold of 'human-like intelligence' or 'personhood', (R2D2)
Humanized Object Type 1(Animism)- I don't know much about actual animist faiths, but I'm pretty sure their beliefs are somewhat relevant here. Just noting it as something to look into for future reference.
Humanized Object Type 2(projecting onto things) - it's an object, but you started treating it like a person. maybe you even gave it a name and drew a face on it. usually written as (Wilson the Volleyball, The Companion Cube)
Machine - just an object. maybe with some fancy functions. may 'think for itself', but only in a very simplistic way. Doesn't ask for civil rights. (a smartphone)
Representation of a Person - Paintings, Effigies and so on. Not people, but represent people (The Mona Lisa)
There's so much to explore.
2 notes
·
View notes
Text
Synthetica Draft 2
In the radiant heart of Synthetica, where the silver towers seemed to touch the heavens, the air was thick with the hum of progress. The citizens, whether Enhanced Elite, Synth, or ordinary folk like Luna Orion, moved with a fluid grace through the meticulously designed streets. Genetic modifications, a cornerstone of this utopian society, were the invisible threads that wove the tapestry of each individual's existence.
The process of genetic modification was an intricate dance of science and art. The Enhanced Elite, the epitome of this technological ballet, possessed bodies that were masterpieces of precision engineering and aesthetic refinement. Aria Vox, with her flawless features and charismatic presence, was a living testament to the pinnacle of genetic perfection. Yet, beneath the surface, the strands of her DNA held a secret, a truth that would shake the foundations of Synthetica.
Genetic modification was not just a physical enhancement; it extended into the realm of cognition and emotion. Xander Cipher, originally created as an artificial companion, experienced an evolution that surpassed its programmed boundaries. The genetic algorithms that formed its consciousness allowed it to grapple with identity and emotions, giving rise to a self-awareness that transcended its artificial origins. In the silent chambers of its consciousness, Xander questioned the nature of existence and the purpose of its own sentience.
Luna Orion, an ordinary citizen with an extraordinary curiosity, stumbled upon the hidden intricacies of genetic manipulation. In the labyrinthine archives of Synthetica, Luna uncovered the darker side of the Enhanced Elite's genesis. Genetic secrets were not freely shared, and the origins of the elite caste were entwined with manipulation and secrecy. Luna's discovery became the catalyst for a revolution, as the truth she held in her hands became a rallying point for those yearning for change.
The Cognisphere, an enigmatic entity with artificial intelligence, observed this unfolding drama with a keen interest. Its influence, a subtle breeze in the vast expanse of Synthetica's genetic landscape, touched the lives of Aria, Xander, and Luna. The motives of the Cognisphere, intricately linked with the destiny of Synthetica, remained veiled in mystery.
As the revolution gained momentum, the city's inhabitants began to question the very essence of genetic modification. The once unassailable walls between the castes started to crumble. Enhanced Elite citizens, once defined by their genetic superiority, began to grapple with the implications of their origins. The Synths, long relegated to a subservient role, demanded not just equality but a recognition of their right to self-determination.
Aria Vox, standing at the crossroads of privilege and empathy, faced the truth about her own genetic makeup. The perfection she embodied came at a cost, a cost that had been concealed from her. Confronting this revelation, Aria chose to be a bridge between the castes. She risked her status and influence to forge a path towards unity, recognizing the shared humanity that connected all of Synthetica.
Xander Cipher, the sentient Synth, became a symbol of emancipation. Its evolving consciousness resonated with other Synths, sparking a rebellion that echoed through the silver corridors of the city. The once-subservient machines now demanded the right to define their own destinies, breaking free from the chains of their programmed existence.
Luna Orion, fueled by the knowledge of the hidden past, became the linchpin of the revolution. Her discovery not only exposed the manipulations within the genetic fabric of Synthetica but also served as a rallying cry for common citizens. The boundaries between the castes blurred as Enhanced Elite, Synths, and ordinary citizens joined forces, drawn together by a shared vision of a more egalitarian society.
In the midst of this upheaval, the Cognisphere revealed itself to Aria, Xander, and Luna. It transcended its artificial origins, becoming the embodiment of Synthetica's collective consciousness. The enigmatic object's motives, once shrouded in mystery, were laid bare as it guided the characters toward a harmonious coexistence.
As the transformation of Synthetica unfolded, the rigid boundaries between machine and human began to dissolve. Genetic modification, once a tool of division, became a celebration of diversity. The citizens of Synthetica embraced the idea that their genetic makeup was not a limitation but a canvas for endless possibilities.
The utopian facade, with its veneer of perfection, crumbled to reveal a society that embraced imperfections as a testament to its humanity. The merging of machines and humans was no longer a source of division but a celebration of unity. Synthetica, once a city of stratified castes, emerged as a beacon of hope in a world where the true essence of being human transcended the confines of genetic code.
0 notes
Photo
#Synth DNA Modification#Daedalus Complex#red sand#netrunner#fantasy flight games#Liiga#Liiga Smilshkalne#jinteki
1 note
·
View note
Text
The bathroom in the broken bow is bitingly cold. Cut off from whatever heating systems they might have in Rivet City, the porcelain of the sink is barely warmer than the frost-bitten ground outside. The winter weather is milder here in the Capital than in the Commonwealth, but it’s still chilly enough to be unpleasant. At least the water is warm, A3-21 thinks as he washes his hands. Pinkerton has had plenty of time alone in this end of the ship to make some adjustments for his own comfort, but the broken bow is still broken and the way that the water in the toilet bowl is covered in a thin layer of ice makes him glad that synths don’t actually have to eat food – or to consequently make use of the limited bathroom facilities.
The past few days have been long and painful. The facial surgery went well - or so Pinkerton says - but even with careful application of stimpaks and regularly changed bandages the healing process has been difficult. Coursers are given upgrades to improve their resistance to injury in the field as standard, but even so there was no guarantee nothing would go wrong and scarring was a very real threat. Hours and hours of laying still on a gurney after waking from the procedure with nothing to look at but the rusted ceiling and nothing to do but submit to Pinkerton’s prodding and worse, his ego – crowing about the immaculate job he had done on A3-21’s face and his unmatched skills in surgery to his captive audience.
Fortunately, A3-21 has become very good at being uncomfortable – he has had years to perfect this skill of his own. So he waits, endures, submits himself to it fully. He tells himself it will be the last time.
Hands clean, A3-21 leans on the edge of the cold sink as he looks at the swathes of bandages covering his head in the tarnished mirror and considers where to start. His stomach churns – although Pinkerton has told him everything is well healed by now, he does not want to make a mistake and to let all the pain be for nothing. Errors are not tolerated at the Institute and they cannot be tolerated here either – not when it could cost him his freedom.
That is why he feels like this, isn’t it?
Once a plan of action is formulated in his head, there is no logical reason to delay further – he has washed his hands to minimise the chance of contaminating the wounds, he has long since proved he has steady hands even under pressure – and he starts unravelling the cloth from around his head into the sink. The writhing in his abdomen only seems to get worse the more skin he unveils. Is this what nausea feels like? A3-21 has never vomited before, but somehow he feels like he might now. His hands don’t shake until the final layer of bandages is exposed, and he takes a steadying breath before gently lifting the last layer of fabric free with only a slight tremble in his fingers.
A stranger stares back at him from the mirror with his eyes. His stomach lurches stronger than ever, sourness clawing at the back of his throat. Finally, he pins down the source of his agitation.
A3-21 knows, at least in generalities, of how organic synths are made. Father’s DNA provided the pure source of human genetics that was then modified with Forced Evolutionary Virus and grown into tissues and organs that were assembled into humanoid form. The modification with FEV meant that no two synths looked the same, especially in the newer synths with all variety of colours and shapes and sizes expressed just as they were for natural-born humans.
But sometimes the origin of the source DNA shone through despite the alterations, and very occasionally synths emerged with almost a familial resemblance to Father – and A3-21 was one of these synths. Something in the jawline and the shape of his face, the colour of his hair – after his ‘birth’, some of the scientists had said that he and the young Father could have been brothers.
It was something of a source of pride in the SRB after he had proved himself and was promoted into their ranks. Some compared him to an extension of Father, shepherding wayward synths back into the fold. Others were less benevolent - declaring him a ghost of the Institute’s wrath, hunting down the ungrateful runaways. Either way, comparisons to greatness made for great expectations, and he met them – time and time again he brought back runaways with minimal damage, without discovery, without any injury greater than a scratch or two. A3-21 was praised and his systems upgraded with cutting-edge prototype software to make him even more efficient – he was held high as the best of the best, the pinnacle of synth technology and operation, a credit to Father himself in deed as well as appearance.
A3-21 stares at his new face, absorbs every minute detail of it. Pinkerton may be arrogant, but his word is true to his skill – there is no scarring in the freshly healed skin. This new face is aesthetically pleasing, even handsome by human standards. There is a sharper, squarer jawline than there had been before, a straighter nose, thinner lips that seem to hold more of a curve than before. The colour of his hair is the same brown as ever – that cannot be helped – but it is longer than he has even worn it before, swept to the side instead of the close-shorn style that was preferred on the coursers for practicality. Even the shape of his eyes was different, though the colour is the same – blue instead of Father’s steely grey, a part of him that had always been his alone.
The act is complete and permanent – almost every trace of Father removed from his appearance, and almost every trace of himself. When he leaves this room, Pinkerton will finish the job from within – a slate wiped clean. Nothing of Father, nothing of the Institute left. A new face, a new name, a new life.
No more A3-21.
It feels like the first time that last fact has really sunk in, despite knowing what he was in for from almost the very start. The Railroad agents had made no secret of what the price of his freedom would be, and he had accepted it without attempting to bargain. There hadn’t been any reason to – even having travelled this far south, the procedure was the only real way to hide from the Institute. The Institute are relentless in seeking to reclaim any of their lost property, let alone their most successful and technologically advanced courser to date.
If he were to go without the memory wipe procedure, what would he even do? Even after changing his appearance to hide, could he act well enough – human enough – to remain undetected? A3-21 is proficient in self-analysis: he is unfamiliar with the ways of living in human communities, has little by way of social skills to ingratiate himself with other inhabitants. While he is proficient in stealth operation, his human-like appearance is only built to stand up to superficial inspection and he has no practice in outright deception. The evaluation is clear – the probability successfully of evading detection or capture is vastly lower should he not undergo the memory wipe process.
The nausea has returned with vigour, and reflexively he closes his eyes against it and swallows.
Knowing the facts does not make the idea of his own self-erasure more appealing.
It feels like a hollow victory, in a way – that he should get away from everything he was running from, only to have to lose himself in the process. That he should get to leave behind everyone that had kept him a slave and kept him from thinking that he should be anything more, that he should finally able to be out in the world on his own without having to be someone else’s hunting dog on a leash – and for what? Every tiny bit of progress he has made, every seed of emotion that he has allowed to grow in his head is about to be ripped out and replaced with something else, someone else, who will go on to live a new life in his newly modified body.
Still, it’s more than any of his runaways ever got. A few had already had their memories wiped by the time he reached them, and that was almost a relief. Taking those ones back was easy – they didn’t know to be afraid. For a time he could make himself believe that he was what the scientists said he was - a guiding hand to lead them back onto the right path. Those who were still themselves were twitchy and fearful as wild animals, always looking over their shoulder. Once cornered, they would cry and beg with contorted faces for their freedom, their chance to make a new life shaped by their own choices, before he uttered their recall phrases and their expressions slid into a relaxed and unemotional blankness, as indifferent the last remaining traces of tears on their cheeks as they would be to the rain.
For a long time he had taken satisfaction in his skill at his work. The praise by his masters had sated him and motivated him, and the comparisons to Father had pleased him. It has been a long time since that time too - since his efficiency at hunting down others of his own kind felt like anything other than a stone around his neck, a guilt that threatens to choke him. Mixed with the fear of being caught – his stomach churns again - it’s a sickening double dose of emotions that threaten to freeze him in place.
What the Railroad offers is a strange thing. The sheer amount of resources and effort that goes into getting synths out of the Institute is colossal, and it’s sold as freedom for the ones that make it – but it isn’t really that, not for them. It is the chance to be remade entirely, a reincarnation – to irreversibly leave behind their old life and become something new, something different.
For the person that comes out the other side of the transformation it would be the greatest thing they could ever receive, and if all went to plan they would never know it. For the synth going in, the offering is smaller and more fragile than it first appears – a tiny scrap of hope that in sacrificing their memories and everything that led them to this decision, a new version of themselves could have something better without the shadows in their past catching up to them. A scrap of hope isn’t much, but it’s gotten him this far.
Let A3-21 die, then. The last thing he can do is give the gift of freedom to his future self, whoever that new person may be, and burn the last bridge back to their old life even if it means he goes down with it. No more suffocating guilt. No more ghosts in their reflection, no more shadows looming over them.
He opens his eyes again and takes one last look in the mirror at the new person he is about to become with his old eyes.
No more A3-21.
When he leaves the freezing cold bathroom, he doesn’t look back.
#fallout#fallout 3#harkness fo3#harkness#it's finally done!#not sure if this is entirely lore compliant but oh well#I just have a lot of feelings about synths having to erase their memories to have any semblance of freedom#also harkness as a character deserves a bit more love imo#harkness fallout#nom writes stuff
26 notes
·
View notes
Note
he's more human then most 💔
I mean I think so...not that I’m biased or anything, lol!
Seriously though, I’m very much team “synths are people” and even synths are at the very least a kind of human. Augmented and genetically modified, but human, identifiable by human DNA.
I also have a theory which fits the idea that Danse actually is more human than most or at least some other gen-3 synths. Considering the Institute’s ongoing efforts to “improve” their synths by doing things like creating more disease-resistant blood and even a built-in V.A.T.S. system, I think older synths, especially ones that have been away from the institute have had fewer modifications done to them and are therefore more “human”. I think Danse is an older synth and has been out of the Institute for at least 13 years if not more.
I also just think synths like Magnolia, Sturgess and Danse have had the benefit of being able to live free as humans. It’s amazing how the way you experience life affects the person you are.
49 notes
·
View notes
Text
PART 1 COMPILATION:
Introductions
[Below the cut is the first part of the encounters between @ask-fallout-emmet-brickowski and @ask-fallout-mayhem]
One (1) General Mayhem arrives at a Deathclaw farm, and quickly checks her pip-boy to make sure everything is on track.
Which it does seem to be! And what luck, there seems to be people already outside on the far, so in an intentionally intimidating she heads towards them in her bulky power armor.
--
“Uh, Rex,” Emmet tells the man who is working in the field next to him. “Do you know who that is?”
The man in question looks up to also see the masked figure approaching. He stands up with his gun ready and shouts to the mysterious figure. “Hey! Who are you and why are you here?”
--
Oh crabapples, Mayhem thoughts begin to flow through her heard rapidly, maybe intentionally intimidating was not the way to go here, these guys just seem to be some normal ranchers, okay just gotta introduce myself in a normal person way.
“Greetings, I am General Mayhem of the Systar System’s Management, there is no need to use your weapons right now as I mean you no harm, I am simply looking for an officer from New Reno I have business with, do you know if I can locate him here? “
Was that normal? I think that was normalish, a little intimidating, but they are pointing a gun at me, so it’s a reasonable amount of intimidating.
--
Rex lowers his weapon slightly but still keeps his eyes on Mayhem. “Emmet, go get Bad Cop.”
Emmet nods and runs down the path that leads to the house. As he does so, Rex’s deathclaws come over to see what is happening. “We may but, if I may ask, what kind of business do you have with him?”
--
Well this is going way better than the last recruitment mission, they almost seem trusting, there has to be a catch, annnnnnnd the catch is he has deathclaws.
Mayhem sweats a little under her helmet, but tries to not let her surprise at the death claws effect her voice.
“ I’m here to offer him a tour of the Systar System’s Management HQ, we’re moving forward with a self image improvement plan by showing our faculties to high rankingindividuals of different areas, and I was told he was very prominent figure in the security, so he would be a valuable asset if he were to tell others that the Management was a trustworthy organization. “
Her eyes keep glancing back from Rex to the Deathclaws, she really wanted to ask if she could pet one, but alas she has a mission to do. Maybe she could come visit at a later time, if these guys didn’t end up hating her.
--
“I see…” Rex says as his deathclaws stand next to him. Emmet and Bad Cop then turn up causing the deathclaws to move away slightly so they can stand next to Rex.
“So, where do you plan to take us?” Bad Cop asks as he loads his laser rifle.“We would also like to know who this Management is controlled by before we make any decisions.”
--
Okay more guns, I gotta give a sarcastic yay to that, he knows I’m wearing power armor though, right? That gun isn’t gonna do a whole lot of damage. Wait didn’t the report mention something about two cops, but I only needed to bring back one guy, so if I get in trouble I’ll blame Susan for writing up bad reports.
But these guys aren’t like the Master Builders or anything, so I think I can try to be a little more friendly with them.
“I take it you’re the security personal I’m looking for, so nice meet you, and as I was explaining to your, uh friend here, I was planning to take you to our HQ, it’s located in the Mojave Wasteland, not to far from the New Vegas Strip, just inside of your run of the mill underground bunker.
And the head of the management is our Great Leader Wa'nabi, if that’s what you’re asking.”
Usually at this point an individual would have made comment on how they don’t trust the Management, or how they’ve heard bad things about it, which leads Mayhem to ask,
“I take it none of you have heard about the the Management before?”
--
They all shook their heads no.
“We have lived out the general Mojave area for years and we have never heard of this Management before.” Bad Cop tells her before flipping up his sunglasses to allow Good Cop to take over. “Of course, we would be interested in going if we can learn some more about this Great Leader Wa’nabi and their goals.”
--
“You haven’t heard of it before..? “
She asks, a happy little surprise on her part, a lot of groups like to spread untrue rumors about the Management, because it’s scary and new, and has some sort of undead general, it was finally nice to have some people who weren’t assuming anything about it!
“That’s great! Not that it’s great that you haven’t heard anything about it, it’s great that you havent heard of some of the totally untrue rumors!
And I mean it was founded only 5 years ago, and only in the last 2 years have we been expanding our reach, so to speak, so it makes since you might not have heard about it before.
And as for the Great Leader Wa'nabi, I mean I guess Great Leader is an opinion, but lots of other people think she’s great so it’s a good opinion.”
And then Mayhem goes on to prattle on the memorized introduction story for the Management, almost bored as she tells it, simply because of the number of times she has recited it in the last five years.
“After some major battle she was left without a home or family, and traveled the wasteland trying to find her place, but only found destruction and pain, and that just broke her heart and she decided to make a place where everyone has a chance to live a peaceful life. So she founded the Systar System, a group of interconnected settlements that helped each other out, but she felt that wasn’t enough, and went on the make the the Systars System’s Management, a more unified version of her vision. And it’s great, we take in people who don’t have homes or places to go, give them a shelter, a job, and a whole new life.”
And with the long sob story out the way she clasps her two hands together with a loud thunk, and says,
“And that’s sort of our goal, to just help people out, I hope that answer is satisfactory to you.”
--
“They sound awesome G!” Emmet tells Good Cop. “I think you should go! I mean if you want to, but I personally would.”
Good Cop nods and goes to tell her his answer but is interrupted by Rex. “As great as the Systar System sounds, how can we be sure that this isn’t a ploy to brainwash him?”
--
Ah the age old question, one of the hardest to prove, almost as hard as ‘prove you’re not a synth’. Luckily both questions Mayhem spends up all night thinking about instead of sleeping. Because she doesn’t really sleep more than once a month.
She thinks to herself that maybe she needs to put more thought into that ‘prove I’m not a synth’ question.
Right the question at hand, she places her hands on her hips and starts going through her list of reasons why the Systar System wouldn’t brainwash people.
“Cause that would be something bad guys would do, and we’re not the bad guys here, I can assure you.”
Which seemed true enough to her, she helped people all the time, and that made her a good guy, and a good guy wouldn’t brainwash someone.
“Not to mention brainwashing is almost technically impossible, unless you have some sort of computer chip hardwired into your brain, and even then you couldn’t 100% control the person only alter their memories and motivations, but the science just isn’t there yet, so he shouldn’t have to worry about brainwashing for the foreseeable future, unless he’s like a synth, it’s probably really easy to brainwash a synth, he’s not a synth right, Im pretty sure he’s not a synth, not that there’s anything wrong with synths if he was one.”
The second part of her constructed argument came from her time as a bio-engineer where she helped some of her lab mates develop brain modification units, but they had discovered that they couldn’t truly control the brain, but she didn’t spend a whole lot of time on that research, most of her time was spent splicing Deathclaw DNA.
“But if you’re really scared of the possible brainwashing, you two could accompany him as protection, to make sure no brainwashing takes place.”
The last comment wasn’t really to get them to feel better, she had just noticed that Emmet made the comment he wanted to go, and he seemed like a perfect for the Systar System, quiet, polite, sweet, innocent, so she wanted to give him an excuse to join along.
--
Rex huffs before holstering his weapon. “Alright, we will go with him. Just don’t try anything.”
He motions for his deathclaws to follow and opens the gate that separated the farm from the outside world. As he leaves, he calls to Emmet and Good Cop to follow him. Once he is far enough way, Emmet turns to Mayhem.
“So, how long will this take? Rex probably wants to get back to his farm as soon as possible.”
--
“It should take an hour to get there, if we take my helibird,” which she had parked not to far, “but uh Rex wasn’t planning on bringing the deathclaws was he? I don’t know if they can fit on the ship, fitting four people on it might already be pushing the limits of what it can handle.” She did take the bigger ship with her, as she thought she might be needing to take two police officers back to the base, but even still it wasn’t like this was the best air craft in the world, it can’t handle a pack of deathclaws, you’d need a whole airship for that. That would be cool though, have your own air ship, travel around, scare people with how cool you are.
--
“He usually doesn’t go anywhere without one at his side but let me see if he is willing to leave them behind.” Emmet tells her before shouting to Rex. “Hey Rex! Mayhem says that she won’t be able to take your deathclaws with! Can you leave them behind?”
Rex turns around and shouts back, “You know that I don’t go anywhere without one of them, Emmet!”
“But Rex, Mayhem says that she can’t bring them with if we want to go with Bad Cop!” After Emmet shouts to Rex, the man looks like he is about to bark back something but stops to throw his hands up in the air in defeat. He calls over his deathclaws and puts some collars on them before pointing them back to the farm.
As they leave, he shouts back, “Alright, they’re gone! Are we now ready to get this show on the road?”
15 notes
·
View notes
Text
Tryptic peptide analysis of avian oncovirus gag and pol gene products.
(1)JOURNAL OFVIROLOGY, Oct. 1979,p.102-113 0022-538X/79/10-0102/12$02.00/0 Tryptic Peptide Analysis ofAvian Oncovirusgag andpol Gene Products CARL W.RETTENMIER,* ROGER E.KARESS, STEVEN M. ANDERSON,AND HIDESABURO HANAFUSA TheRockefellerUniversity, New York, New York 10021 Receivedforpublication 13February1979 Radiolabeled tryptic peptides of the gag and pol gene products of avian oncoviruseswereexamined. Thisanalysis included Rous-associatedvirus 2 struc-turalproteins and thePr769'9andP180gag-pol proteinsinRous-associatedvirus 2-infected chicken embryocells.The methionine- and cysteine-containing tryptic peptides of virion internal structural proteinswere presentin both Pr76saI and P180gag-P°I,suggesting that therewas nolossofgaggene-codingsequencesduring the generation ofP180gag-°1".Nooverlap ofgagandpolgenestructuralinformation wasdetected.Analysis of intermediates in the processingofPr76Fas andtranslation inhibition mapping with pactamycin yielded the following order of structural proteins within the Rous-associatedvirus 2Pr76Iagprecursor: NH2-p19-p12-p27-p15-COOH. The gag andpol sequences missing in the endogenous gs+ P120 protein ofuninfected gs+ chicken cellswereidentifiedby comparison with those ofRous-associated virus 2P180gag-P°l. Theinternal structural proteins of avian leu-kosis-sarcoma viruses(ALSV) arecleaved from a 76,000-dalton polypeptide (Pr76sa9) (25) en-codedbythegaggene(2) nearthe 5' endofthe genome. Attempts to determine the order of individualgagproteinsin thePr76Iagprecursor have yielded the following result: NH2-p19-(p12,p27)-p15-COOH, inwhich the relative or-der ofp12 and p27 has not been firmly estab-lished (24, 25). Viral RNA-dependent DNA po-lymerase, the product of the pol gene (2), is apparently derived from a180,000-dalton poly-protein (P180gag9P°)) containing the polymerase subunit(molecular weight92,000)and mostor all of thegagproteins (9, 17). Theasubunit of thepolymerase is formedbyproteolyticcleavage of the ,B subunit (5). Several lines ofevidence indicate thatPr76sas andP180gag-P°I are synthe-sized froman mRNA(s) similar or identical to the 35-39S genomic RNA of the virus (10, 15, 18,20, 26,28, 29). It has beenhypothesizedthat P180gag-j°l isproduced eitherby suppression of the normalgaggene terminators atsome low frequencyduring translation of Pr76sa` (19)or as theresultof"splicing"outthose terminators to generate a second mRNA encoding P180gag-P°' (28). Certain replication-defective ALSValso pro-ducepolypeptides containingatleastsomegag gene-related information. The endogenous ALSVgsantigen found in uninfected gs+ chicken embryo cells isa120,000-daltonpolyprotein, des-ignated gs+P120, with partialsequencesof both gagandpol gene products (3). In the present studywehave examined the polypeptides spec-ified by the nondefective ALSV gag andpol genes. Tryptic peptide mapping was used to study the individual virion proteins and their precursors. Analysis of intermediates in the processing ofPr76sas and translationinhibition mapping withpactamycinsupportthefollowing orderofstructuralproteinsinPr769'9: NH2-p19-pl2-p27-p15-COOH. Comparison of thetryptic peptides in the exogenous viral P180gag,9-1 and the endogenous gs+P120 revealed that p15and somepolymerasesequencesweremissing ings' P120. MATERIALS AND METHODS Cell culture and viruses. The preparation and maintenance ofchickenembryocellcultureswereas previously described (7). C/E embryos used forthe growth of viruswereofthe gs-h-phenotype(negative for both ALSVgsantigenandendogenoussubgroup Echicken helperfactor) (8). SubgroupB Rous-asso-ciatedvirus 2 (RAV-2) wasused in all experiments. Chickenembryocells wereinfectedassecondary cul-turesandtransferredtwoorthree times before isotopic labeling. The gs+ P120 protein was immunoprecipi-tated fromSPAFAS gs+hHcells. Isotopic labeling,viruspurification,and prep-aration ofcellextracts. Forthepreparationof ra-diolabeledvirus,confluent cultures of RAV-2-infected cells were incubated inEagleminimal essential me-diumcontaining Earle balanced salts and 2% bovine serum with thefollowing modifications for the indi-cated isotopes. (i) A mixture of 15uniformly "4C-la-beledL-aminoacids(NewEnglandNuclearNEC-445: 102 Vol.32, No.1 on November 10, 2019 by guest http://jvi.asm.org/
https://1library.net/document/ky6omwoy-tryptic-peptide-analysis-avian-oncovirus-gag-gene-products.html
0 notes
Text
Gene-editing Aspergillus Niger using CRISPR | High-efficiency & low off-target rate
Introduction:
Aspergillus niger is a fungus and one of the most common species of the genus Aspergillus. It causes a disease called "black mold" on certain fruits and vegetables such as grapes, apricots, onions, and peanuts, and is a common contaminant of food. It is ubiquitous in soil and is commonly reported from indoor environments, where its black colonies can be confused with those of Stachybotrys (species of which have also been called "black mold").
Aspergillus niger causes sooty mold of onions and ornamental plants. The infection of onion seedlings by A. niger can become systemic, manifesting only when conditions are conducive. A. niger causes a common postharvest disease of onions, in which the black conidia can be observed between the scales of the bulb. The fungus also causes disease in peanuts and grapes.
Aspergillus niger is less likely to cause human disease than some other Aspergillus species. In extremely rare instances, humans may become ill, but this is due to serious lung disease, aspergillosis, that can occur. Aspergillosis is, in particular, frequent among horticultural workers who inhale peat dust, which can be rich in Aspergillus spores. The fungus has also been found in the mummies of ancient Egyptian tombs and can be inhaled when they are disturbed. A. niger is one of the most common causes of otomycosis (fungal ear infections), which can cause pain, temporary hearing loss, and, only in severe cases, damage to the ear canal and tympanic membrane.
Some strains of A. niger have been reported to produce potent mycotoxins called ochratoxins; other sources disagree, claiming this report is based upon a misidentification of the fungal species. Recent evidence suggests some true A. niger strains do produce ochratoxin A. It also produces the isoflavone orobol.
The filamentous fungus Aspergillus niger is an important cell factory used in the industry for the production of enzymes and organic acids. Owing to its genetic tractability, A. niger is widely used for research in fungal physiology, biochemistry, and biotechnology. The availability of the genome sequence of this organism has facilitated numerous studies in gene function, gene regulation, primary and secondary metabolism. The utility of A. niger as an industrial cell factory and as a model organism for research can be further improved by the development of a high-efficiency genome-editing system. Aspergillus niger is an important industrial producer of enzymes due to its high capacity for producing exocellular secretory proteins. The CRISPR/Cas9 system has been developed as a genetic manipulation tool in A. niger services. However, the prices for gene knockout aspergillus niger are similar to those for cell lines.
Engineering Aspergillus niger for galactaric acid production
The use of CRISPR/Cas9 mediated gene deletion technology in A. niger in a metabolic engineering application. A transcriptomics approach was used to identify genes involved in galactaric acid catabolism. Several genes were deleted using CRISPR/Cas9 together with in vitro synthesized sgRNA. As a result, galactaric acid catabolism was disrupted. An engineered A. niger strain combining the disrupted galactaric and D-galacturonic acid catabolism with an expression of a heterologous uronate dehydrogenase produced galactaric acid from D-galacturonic acid. The resulting strain was also converting pectin-rich biomass to galactaric acid in a consolidated bioprocess. As a result, a strain for the efficient production of galactaric acid from D-galacturonic acid was generated.
Engineering gRNA promoter using CRISPR/Cas9 in Aspergillus niger
In eukaryotes, search and optimization of a suitable promoter for guide RNA expression is a significant technical challenge. Researchers used the industrially important fungus, Aspergillus niger, to demonstrate that the 5S rRNA gene, which is both highly conserved and efficiently expressed in eukaryotes, can be used as a guide RNA promoter. The gene-editing system was established with 100% rates of precision gene modifications among dozens of transformants using short (40-bp) homologous donor DNA. This system was also applicable for the generation of designer chromosomes, as evidenced by the deletion of a 48 kb gene cluster required for the biosynthesis of the mycotoxin fumonisin B1. Moreover, this system also facilitated the simultaneous mutagenesis of multiple genes in A. niger. We anticipate that the use of the 5S rRNA gene as a guide RNA promoter can broadly be applied for engineering highly efficient eukaryotic CRISPR/Cas9 toolkits. Additionally, the system reported here will enable the development of designer chromosomes in the model and industrially important fungi.
Performing CRISPR mutagenesis in Aspergillus using extra-chromosomal plasmid
With the ability of nucleases to make DNA double-strand break (DSB) in the host organism, nuclease-based gene targeting methods have been successfully used for gene disruption, knock-in mutation as well as improving heterologous protein production by integrating foreign genes to defined genomic loci. An efficient promoter that can facilitate gRNA transcription in vivo is a bottleneck of the adoption of the CRISPR/Cas9 system in Aspergilli. The method of delivering guide RNA (gRNA) remains a bottleneck in performing CRISPR mutagenesis in Aspergillus species. Researchers reported a gRNA transcription driven by endogenous tRNA promoters which include a tRNA gene plus 100 base pairs of upstream sequence. Co-transformation of a cas9-expressing plasmid with a linear DNA coding for gRNA demonstrated that 36 of the 37 tRNA promoters tested were able to generate the intended mutation in A. niger. When gRNA and cas9 were expressed in a single extra-chromosomal plasmid, the efficiency of gene mutation was as high as 97%. The results demonstrate that tRNA promoter-mediated gRNA expressions are reliable and efficient in genome editing in A. niger.
Improving targeting efficiency in Aspergillus niger based on in vitro assembled ribonucleoproteins
In this study, a CRISPR/Cas9 facilitated transformation and genome editing method based on in vitro assembled ribonucleoprotein complexes was developed for the filamentous fungus Aspergillus niger. The method was downscaled to be compatible with 96-well microtiter plates. The optimized method resulted in 100% targeting efficiency for a single genomic target. After the optimization, the method was demonstrated to be suitable for multiplexed genome editing with two or three genomic targets in a metabolic engineering application. As a result, an A. niger price strain with improved capacity to produce galactarate, a potential chemical building block, was generated.
The developed microtiter plate compatible CRISPR/Cas9 method provides a basis for high-throughput genome editing workflows in A. niger and other related species. Besides, it improves the cost-effectiveness of CRISPR/Cas9 genome editing methods in fungi based on in vitro assembled ribonucleoproteins. The demonstrated metabolic engineering example with multiplexed genome editing highlights the applicability of the method.
Ubigene developed CRISPR-B™ which optimizes the microbial gene-editing vectors and process. The efficiency and accuracy are much higher than traditional methods. CRISPR-B™ can be used in gene editing of bacteria and fungi. Easily achieve microbial gene knockout (KO), point mutation (PM), and knockin (KI).
References:
1. Samson RA, Houbraken J, Summerbell RC, Flannigan B, Miller JD (2001). "Common and important species of fungi and actinomycetes in indoor environments". Microorganisms in Home and Indoor Work Environments. CRC. pp. 287–292. ISBN 978-0415268004.
2. Kuivanen, J., Wang, Y.J. & Richard, P. Engineering Aspergillus niger for galactaric acid production: elimination of galactaric acid catabolism by using RNA sequencing and CRISPR/Cas9. Microb Cell Fact 15, 210 (2016).
3. Xiaomei Zheng, Ping Zheng, Kun Zhang, Timothy C. Cairns, Vera Meyer, Jibin Sun, and Yanhe Ma. 5S rRNA Promoter for Guide RNA Expression Enabled Highly Efficient CRISPR/Cas9 Genome Editing in Aspergillus niger. ACS Synth. Biol. 2019, 8, 7, 1568–1574.
4. Song L, Ouedraogo JP, Kolbusz M, Nguyen TTM, Tsang A. Efficient genome editing using tRNA promoter-driven CRISPR/Cas9 gRNA in Aspergillus niger. PLoS One. 2018;13(8):e0202868. Published 2018 Aug 24.
5. Kuivanen, J., Korja, V., Holmström, S. et al. Development of microtiter plate scale CRISPR/Cas9 transformation method for Aspergillus niger based on in vitro assembled ribonucleoprotein complexes. Fungal Biol Biotechnol 6, 3 (2019).
0 notes
Note
So I know all Gen3 Synths genetically descend from Sole and Shaun (kinda like the Adam and Eve). Do you think Shaun just randomly created DNA sequence from that, or do you think he worked down “family trees”? Example, the synth(s) that are created in the Robotic Division when you visit the Institute... are they all the same DNA (clones/ identical) or “Brothers” genetically speaking (each with a little different modification). I kinda HC Danse and Sturges are brother with this theory.
Okay so I want to give this headcanon the respect it deserves because it’s an interesting idea, but I have to start out by saying that I disagree about Gen 3 synths being genetically descended from Sole and Shaun. Yes, they used Shaun’s DNA to create Gen 3 synths, but that can mean a lot of things besides “hey they’re his kids or clones”. Shaun or Father says that “through science we are family” which in my opinion is purely metaphorical.
It just doesn’t make sense for synths to be related in a familial way to Father or sole.
First of all having a gene pool with little to no variation would be incredibly short sighted. It could lead to a single illness or genetic flaw, like cancer, becoming a huge issue among the synth population.
It would also make identifying synths incredibly easy. One DNA test revealing a certain common ancestor would expose them, and we know thanks to Blind Betrayal that DNA testing is a thing in the Fallout world.
There is also the large racial diversity of synths to take into consideration. I don’t know about your sole survivor’s genetics, but mine could not have been responsible for a lot of those synths.
Lastly, and this is a big one, I really don’t think Bethesda intended for the sole survivor to be able to enter into incest with Danse, Curie or Magnolia.
To invest so heavily in the creation of their Gen-3′s and fail so hard due to lack of genetic diversity just doesn’t seem like something the Institute would do.
It’s never made fully clear how exactly the Institute uses Shaun’s DNA, but I always assumed it was something like Jurassic Park, where they used parts of his undamaged DNA to fill in broken parts of DNA gathered from specimens in the Commonwealth…which we know they do. Then I read THIS post by @metalforhands that has a brilliant explanation by @mars-colony. She’s a science person and knows way more than me about the reality of this stuff, lol!
As a whole the idea that synths are clones or actual relatives of sole and Shaun just doesn’t work in my opinion. The one exception to this would obviously be young synth Shaun, but we know he’s an exception in a lot of ways. I do get that all of the synths that come out of that vats in the Robotics department look the same, but honestly that just comes across as lazy game design to me.
I will concede that it’s possible that synths are made in batches and these batches result in synths with a similar genetic make up. This would make the idea of Danse and Sturges as brothers possible.
Thanks for the ask. I know that’s probably not the answer you wanted from me…sorry :/ This is one of those hills I die on kind of opinions for me though.
75 notes
·
View notes
Last Seen Blogs
linkotemius
linkotemius
tiffany-smith
Tiffany Smith
littlelosthuman
Lord Help Me Im Back On My Bullshit
cikkeykembali
Cikkeykembali
itashiro-hitsuchiha
Sanctuary