Скачать или смотреть “CRISPR” as molecular manipulator is a rapidly-expanding toolbox. Here’s a flavoring...

CRISPR/Cas is a powerful gene editing tool along with other things, it uses a guide RNA that binds to a CAS protein, directs that CAS protein to a DNA, and then the CAS protein takes over. You can use different CAS proteins to have different effects.

MUCH more here: https://bit.ly/crisprcasscience &    • CRISPR/Cas Science - Gene editing & beyond...  

Conventional CRISPR/Case uses Cas9, which cuts both strands of the DNA and leaves the cell to fix things. If you don't provide an alternative, you typically get knocked out where the gene is basically inactivated. If you give it an alternative, you can have something called homology directed repair that allows for gene editing. However, both of these strategies have issues and they're not very efficient.

More recent strategies use nickase Cas9 (nCas) proteins, where they basically make a cut to a single strand and then are associated with an enzyme that makes a modification to the DNA.

In base editing (BE), you're changing a single nucleotide to make a point change, typically with a damages. With prime editing (PE), you use reverse transcriptase and to provide a template to make larger changes. Unlike in the case of your conventional like gene editing through homology to repair. However, this template is coming from the guide RNA itself, not from an external piece of DNA.

If you want non-permanent changes, you can also use catalytically-dead (dCas) proteins attached to things like transcription factors or chromatin modifiers that allow you to temporarily change expression. You want to change what sequence is targeted, you change the guide RNA, and if you want to change what happens, you change the CAS protein. In addition to gene editing, you can also do transcriptional regulation.

You can target RNA with specific Cases, and you can do things like use Crispr as a diagnostic tool.

There are, however, challenges. These include trying to make edits that are accurate and efficient, targeting to every sequence that we want. So overcoming some of those issues with needing to be next to the Pam, delivering the machinery, evading the immune system. And so there's lots of different strategies and I'll provide you with links to resources that do a great job explaining all this.

There's a lot more coming down the pipeline. And so I encourage you to check out those free papers. None of this could have happened, however, without the original basic research
                             
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