AUTHOR=Jiang Hua , Duan Kun , Han Xu , Wang Jun , Liu Xiao , Yan Maoxiao , Wang Yunxiu , Liu Hongyan , Shi Huiling , Gao Xiaoqing , Ouyang Chuan , Fu Xue , Zhang Xinxin , Liu Chao TITLE=Detection of Mitochondrial Mutations Through Isothermal Nucleic Acid Amplification Coupled With Clustered Regularly Interspaced Short Palindromic Repeat-Associated Endonuclease Cas13a JOURNAL=Frontiers in Genetics VOLUME=11 YEAR=2021 URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2020.622671 DOI=10.3389/fgene.2020.622671 ISSN=1664-8021 ABSTRACT=
The clustered regularly interspaced short palindromic repeat (CRISPR)-associated endonuclease Cas13a can specifically bind and cleave RNA. After nucleic acid pre-amplification, bacterial Cas13a has been used to detect genetic mutations. In our study, using a transcription-mediated amplification together with Cas13a, we can isothermally amplify and detect mitochondrial point mutations under non-denaturing conditions from human genomic DNA. Unlike previous reports, we prepared CRISPR DNA with T7 promoter sequences and generated CRISPR RNA