Crispr-Cpf1 HELP PLEASE!

So, news around town is that Crispr-Cpf1 works better than Crispr-Cas9 systems, as the protein is smaller and can fit in more places, it’s smaller not only in size, but in base pairs, so it can fit in viral vectors better, and doesn’t require all that Crispr-Cas9 requires. Even the PAM is more desirable.

Would it make sense or be feasible, to design a phage for Prevotella (has cpf1) that contains the virus of choice, and allowing the virus to make its own gRNA and molecular scissors, separating that plasmid and using that to turn into a AAV virus for therapy?

For instance, can one use BX795 or another inhibitor to lower cellular immunity, say a gel of Bx795 in the vagina and applying a Crispr gel, allowing Crispr to do its job without the body trying to defend against its Cas9/Cpf2 systems?

Comments

  • This is a shotgun blast of concepts. Let's try to work through them all.

    A lenti virus can easily handle a crispr system. I think our latest construct has two know outs and isn't too big to mess anything up.

    If you want to make a virus for human application, you need to grow that virus in human cells. It just works better. You can insert your lenti plasmids (usually a 3 part system) into you mammalian cells with electroporation or better yet, lipofectamine, and then they build themselves.
    You can build your plasmids in simple ecoli, isolate, and then bake in mammalian cells. We're having good luck with the Vero line, but CHO is also good.

    Your last paragraph needs clarification. What is "Crispr gel"? Also, the targeting and therapy outline seems highly specific. Have you looked in to what Andreas Stuermer is doing with glyD knockout herpes?

    Remember, just cause a tool exists, doesn't mean it need to be used.

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