Gene editing with CRISPR-Cas12a guides possessing ribose ... - Nature
Nov 15, 2021 · Here we chemically modify ribonucleotides of the As Cas12a CRISPR RNA 5′ handle, a pseudoknot structure that mediates binding to Cas12a. Gene editing in human cells …
Biology and applications of CRISPR–Cas12 and transposon
Dec 4, 2024 · Cas12a and Cas12i use a catalytic site that resides in the WED domain to cleave their pre-crRNA upstream of each crRNA pseudoknot in a metal-independent manner 41, 62.
Binding to the conserved and stably folded guide RNA pseudoknot …
May 1, 2023 · Here, we propose that Cas12a RNP assembly with its cognate CRISPR RNA (crRNA) guide instead proceeds primarily through Cas12a conformational changes during …
Optimized protocols for the characterization of Cas12a activities
Cas12a binds to a cognate RNA called the CRISPR RNA (crRNA), specifically by recognizing the 5′ end of the crRNA that possesses a pseudoknot structure to form a binary complex (Yamano …
Kinetic dissection of pre-crRNA binding and processing by CRISPR-Cas12a …
When the RNA binds Cas12a to form a RNA-protein complex (RNP), the pseudoknot binds into a cleft between domains of Cas12a and is further stabilized by coordinating two hydrated Mg 2+ …
Regulation of the CRISPR-Cas12a system by methylation and …
These findings show that the deactivation of CRISPR-Cas12a relies on the synergetic effect of multiple m6A modifications on the pseudoknot structure. Notably, m6A modifications at the …
Structural Basis for Guide RNA Processing and Seed-Dependent …
Apr 20, 2017 · The structure of a binary complex of Lachnospiraceae bacterium Cas12a (LbCas12a) bound to a crRNA has revealed that the repeat-derived part of the crRNA forms a …
Good guide, bad guide: spacer sequence-dependent cleavage …
Apr 4, 2020 · (A) Cas12a binds the repeat of the pre-crRNA, which forms a typical pseudoknot structure (blue). It recognizes a TTTV PAM (Yellow) and forms an R-loop with the spacer part …
Gene editing with CRISPR-Cas12a guides possessing ribose …
Nov 15, 2021 · Here we chemically modify ribonucleotides of the As Cas12a CRISPR RNA 5′ handle, a pseudoknot structure that mediates binding to Cas12a. Gene editing in human cells …
Gene editing with CRISPR-Cas12a guides possessing ribose
Nov 15, 2021 · Here we chemically modify ribonucleotides of the AsCas12a CRISPR RNA 5' handle, a pseudoknot structure that mediates binding to Cas12a. Gene editing in human cells …