Influence of N1-Methylpseudouridine in Guide RNAs on CRISPR/Cas9 Activity.

ABSTRACT: At present, there are many strategies to improve the activity of CRISPR/Cas9. A well-known and effective approach is guide RNA modification. Many chemical guide RNA modifications have been studied, whereas naturally occurring RNA modifications are largely unexplored. N1-methylpseudouridine (m1Ψ) is an RNA base modification widely used in mRNA therapy, and it holds great promise for application in genome editing systems. The present study focuses on investigating the effect of N1-methylpseudouridine on the functioning of CRISPR/Cas9. In vitro cleavage assays helped determine the level of m1Ψ guide RNA modification that is sufficient to cleave the target substrate. By analyzing FAM-labeled dsDNA substrate cleavage, we calculated the kinetic parameters and the specificity scores of modified g

SUBMITTER: Prokhorova D 

PROVIDER: S-EPMC10707292 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature