期刊
TALANTA
卷 210, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.talanta.2019.120613
关键词
CRISPR/Cas9; CRISPR/Cas12a; Biallelic mutants; Mutants screening
资金
- Natural Science Foundation of China, China [81873958]
- State Key Laboratory of Respiratory Diseases Open Project, Shenzhen, China [SKLRD-OP-201919]
- Shenzhen Scientific and Technological Foundation, Shenzhen, China [JCYJ20170412151620658, JCYJ20170307095003051, JCYJ20170307095303424]
- Special Support Funds of Shenzhen for introduced High Level Medical Team, Translational Medicine of Biochip in Clinical Laboratory, Shenzhen, China [SZSM201412005]
CRISPR/Cas9 is a robust tool to manipulate genes in a wide range of species. Although several methods are introduced to identify the CRISPR/Cas9-induced mutations, they are labor-intensive, costly, and not easy to use or were sequence-limited. Moreover, few of them could identify the biallelic mutants that are the desired outcomes of targeted mutagenesis. Recently, a CRISPR/Cas12a-mediated biosensing platform was developed to detect nucleic acids based on the collateral DNA cleavage activity of Cas12a; it was highly sensitive, specific, rapid, and cost-efficient for genotyping, mutation detection, and single nucleotide polymorphism (SNP) identification, thereby deeming it as an innovative method for screening the CRISPR/Cas9-induced biallelic mutants. Thus, the CRISPR/Cas12a-based biosensing platform has been successfully utilized for screening 23 CRISPR/Cas9-induced biallelic mutants in Thp-1 cells, which were also confirmed by direct sequencing and ELISA. The precision and efficiency of CRISPR/Cas12a-based biosensing platform make it a promising tool for screening of CRISPR/Cas9-induced biallelic mutants in the future.
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