Cas12a-based one-pot SNP detection with high accuracy.

ABSTRACT: CRISPR-Cas12a based one-pot detection system has been used in nucleic acid detection and diagnosis. However, it is not sensitive enough to distinguish single nucleotide polymorphisms (SNP), which has greatly restricted its application. To overcome these limitations, we engineered a LbCas12a variant with enhanced sensitivity against SNP, named seCas12a (sensitive Cas12a). SeCas12a-based one-pot SNP detection system is a versatile platform that could use both canonical and non-canonical PAM, and was almost not limited by mutation types to distinguish SNPs located between position 1 to 17. The use of truncated crRNA further improved SNP specificity of seCas12a. Mechanistically, we found only when the cis-cleavage was at low level between 0.01min^-1 and 0.0006 min^-1

SUBMITTER: Zhang HX 

PROVIDER: S-EPMC10134196 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature