RE: [CRISPR 101] What is CRISPR?
Good to see introductions to CRISPR on here, and not the fear-mongering kind!
CRISPR/Cas9 is but one of the many CRISPR effectors though, with Cpf1 making much progress to becoming a more precise genome engineering tool. Though, this precision does come with a trade-off of overall enzymatic activity, and less ability to displace nucleosomes the way Cas9 does.
The part I find most remarkable about this is that the "scanning" of Cas9 (or Cpf1) for the proper target that matches the gRNA, target recognition, and hydrolysis of target DNA, is all ATP and NADH/NADPH independent. That such robust activity can occur without additional energy input is quite something. Once assembled, this system works remarkably, despite the chance of off-targets; but then again there are the new hyper-accurate Cas9 variants, or Cpf1 if specificity is an issue.
Looking forward to the next in the series, and if you have any questions, feel free to hit me up; I work with these tools and would relish the opportunity to blab on about my work!
Cheers!
Thank you for your comment! I also work in the CRISPR field so i'm definitely more excited than concerned about the technology :) I'm glad that you like the it, i'll be happy to have your feedback on future publications since i don't have a lot of experience in the writing of this kind of articles.
You raised some pretty interesting points here. I am not a Cpf1 expert but i think that a very interesting (and relatively unexplored) feature of it is that it produces a staggered cut, which could lead to different repair outcomes compared to Cas9. I think that in the long term we will see these different enzymes fit into a molecular toolbox that could be adapted to specific applications (longer or shorter PAM, cutting activity...). If you are interested in accuracy, here is a promising report about the Cas9 from Nesseiria Meningititis (still not peer-reviewed thought):
https://www.biorxiv.org/content/early/2017/08/04/172650
The energy question is also very interesting, I think the fact that DNA could be cleaved simply using the free energy stocked in the molecule must have been a critical reason of why it has become the support of heredity at the early stages of evolution, but this is just speculations at that point :)