Cas12 and Cas13 in Genome Editing: Beyond CRISPR-Cas9

References/Resources: https://www.patreon.com/the_Crux

All videos on Genetic Engineering: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf

This video broadly discusses Cas9, Cas12, Cas13, Cas12f, and Cas3 enzymes. Essentially, we discuss alternative Cas enzymes used in gene editing and genome engineering. Our discussion begins with the classic SpCas9 system and discuss its variants, PAM requirements, off-target effects, and compare different engineered Cas9 high-fidelity variants including HyperAccurate Cas9, xCas9, and PAM-less SpRY-Cas9. We also compare naturally occurring Cas9 orthologs such as SaCas9, SauriCas9, and PsCas9, highlighting differences in size, specificity, and DNA cleavage properties. In Cas12 family, we discuss Cas12a/Cpf1, Cas12b, Cas12e/CasX, and CasY, and how their T-rich PAMs, RuvC-mediated cleavage, sticky-end DNA cuts, are useful for targeting AT-rich genomic regions and 3′ UTRs. In contrast, Cas13 enzymes target single-stranded RNA instead of DNA. They have a compact gRNA, tend to require a protospacer flanking sequences (PFS), and exhibit a phenomenon of collateral RNA cleavage. Finally, we discuss Cas12f and Cas3 enzymes. Cas12f is a compact nuclease capable of single-stranded DNA cleavage and collateral DNA damage. The Cascade-Cas3 system is a processive DNA-degrading CRISPR complex with helicase and ATPase activity, capable of shredding DNA over long distances. The video concludes with other recent advances in CRISPR engineering, including DNA-guided RNA-targeting Cas12 variants and the PAM-independent TIGR-Tas system developed by Feng Zhang’s group at MIT.

00:00 Agenda
00:44 SpCas9 and variants
03:28 Cas12 enzymes
05:10 Cas13 enzymes
06:33 Collateral cleavage
07:11 Cas12f enzyme
07:47 Cas3-Cascade
08:35 TIGR-Tas system

CRISPR-Cas9 in genome editing: https://youtu.be/7ESZTE6rjLI

References/Resources: https://www.patreon.com/the_Crux

*Must watch playlists*
All videos on Genetic Engineering: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf
All videos on DNA Replication: https://youtube.com/playlist?list=PL0Ymnd-zt4IgLefDaeqicJbCBnbLqnDJ7
All videos on Transcription: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij2VcAOHNUweftSElVsc-MX
All videos on Translation: https://www.youtube.com/playlist?list=PL0Ymnd-zt4IjAAAMlPLDIXbs692Yp14xa

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Видео Cas12 and Cas13 in Genome Editing: Beyond CRISPR-Cas9 канала theCrux