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Is it possible to transfect the full purified DNA/RNA of a virus into a cell culture and obtain working infectious viral particles? #60

Open cirosantilli opened 4 years ago

cirosantilli commented 4 years ago

Coming from a non-biology background, my gut feeling is yes, sine this is basically what viruses themselves to. If that is the case, could anyone point to papers that have achieved that to confirm?

The difference between what I'm asking and what viruses do is that viruses have to have the viral particle fully assembled to be infectious, and I'm thinking instead if it would be possible to bootstrap a virus from the purified DNA/RNA without those already assembled particles.

I'm interested both in bacteria, e.g. with a well studied bacteriophage, and in eukariotes.

The main motivation of this question is to decide if we had a cheap benchtop de novo DNA/RNA synthesis machine, how easy would it be for someone to download the sequence of a deadly virus and produce it, a debate which is very similar to the control of 3D printed guns.

For example, from this molecular description of COVID-19, it seems that once the viral RNA gets into the cell, it goes straight into the host Rybossome without needing any other proteins: https://www.youtube.com/watch?v=8_bOhZd6ieM so if we could synthesize RNA or (or DNA + in-vitro transcription if that exists), then that might be enough.

The final step would be how to get the RNA into the culture without the viral proteins, which is basically what the viral proteins do automatically, so do we have efficient methods for that, to deliver a 30kb RNA into a cell:

Alernatively, we could try to just do everything in a cell-free, which overcomes the hurdle of entering the cell in the first place. However, at least for COVID-19 it steals a lipid layer from the host cell, and it is not infectious without that layer, so maybe this is not the best approach.

Yet alternatively, if the following viral engineering/trasnfection mechanisms are off-the-shelf enough, they would also work:

cirosantilli commented 4 years ago

In https://www.nature.com/articles/s41586-020-2294-9 they've done it seems, functional COVID viral particles from synthetic DNA.