Virus modification

[v30000d]

Hello! I want to know how a biological virus can be modified. Does it require observation through an electron microscope? I searched for step by step instructions but could not find

[iNow]

Given your current lack of even basic knowledge, you simply shouldn’t even try. 

[v30000d]

I want to get a general idea. I have no money for real experience and will not appear in the next 5 years.

Edited April 12, 2021 by v30000d

[CharonY]

I think it would help if you get a basic textbook on viral genetics. But to answer your question, no generally it does not involve looking at them using EM.

[fredreload]

Hmm, the idea is you need to get the genetic material into a nucleus. A while ago there was this building life from scratch where they create a goat virus DNA from scratch(Yes a goat virus for safety precautions so if it gets out of the lab it would not infect humans). I am not sure their method of creation, I guess it is probably something similar to 3D printing DNA, once you have the DNA, insert/swap it into a nucleus of any cell and it should work.

P.S. A virus does have a nucleus right? Wouldn't know, but you need to get the DNA into a cell to start producing proteins from the ribosomes.

[CharonY]

12 hours ago, fredreload said:

P.S. A virus does have a nucleus right?

No. The method used is typically solid-phase synthesis. But no, if you have no training or background one should not attempt it.

[fredreload]

Alright, I got a better idea. This is a virus, shown on this site(https://ysjournal.com/bacteriophages-the-case-for-viruses-in-treatment-development/).

You extract the DNA, modify it with a crispr, and package it into another virus, and place it onto a cell for insertion of genetic material and replication. Simple right? Just swap the DNA genetic material.

[CharonY]

These are bacteriophages, i.e. they only infect bacteria and there is no need to use CRISPR. They have been used traditionally as a cloning vector since they do not care too much about what is packaged in their heads and can therefore be used for specialized transduction. Nowadays there are versatile in vitro systems that utilize viral recombination reactions which makes the virus itself redundant and can be used to e.g. create mammalian expression vectors and other purposes outside of bacteria.

[fredreload]

17 hours ago, CharonY said:

e.g. create mammalian expression vectors and other purposes outside of bacteria.

Hmm, the problem with virus introduced expression vectors have always been that it cannot be integrated with the host genome. Or rather, the virus cannot insert the expression vector at a correct spot in the host genome and that is why crispr is introduced. The mammalian expression has always been, after insertion, expressing itself independently from the host genome. Well, I wouldn't know if there is a way to insert the viral vector at a correct place, but then that would mean we would not need crispr do we?

[CharonY]

4 hours ago, fredreload said:

Hmm, the problem with virus introduced expression vectors have always been that it cannot be integrated with the host genome. Or rather, the virus cannot insert the expression vector at a correct spot in the host genome and that is why crispr is introduced.

You were talking about phages. They only work with bacteria and it is fairly to get a site specific construct. Also expression systems typically do not require site-specific insertion into the host genome. The goal there is not to modify the genome, but to express specific proteins (or RNA). 

Site-specific mutations of eukaryotes are an entirely different thing, but again, you would not use bacteriophages for that one. I mean, if you are actually interested there are a whole bunch of books (including open source textbooks) you could read on that topic.