染色体易位
生物
基因组编辑
清脆的
核酸酶
Cas9
转录激活物样效应核酸酶
染色体工程
遗传学
锌指核酸酶
计算生物学
DNA
基因
作者
Jiyeon Kweon,Hye‐Yeon Hwang,Haesun Ryu,An-Hee Jang,Daesik Kim,Yongsub Kim
标识
DOI:10.1016/j.ymthe.2022.09.008
摘要
A variety of cancers have been found to have chromosomal rearrangements, and the genomic abnormalities often induced expression of fusion oncogenes. To date, a pair of engineered nucleases including ZFNs, TALENs, and CRISPR-Cas9 nucleases have been used to generate chromosomal rearrangement in living cells and organisms for disease modeling. However, these methods induce unwanted indel mutations at the DNA break junctions, resulting in incomplete disease modeling. Here, we developed prime editor nuclease-mediated translocation and inversion (PETI), a method for programmable chromosomal translocation and inversion using prime editor 2 nuclease (PE2 nuclease) and paired pegRNA. Using PETI method, we successfully introduced DNA recombination in episomal fluorescence reporters as well as precise chromosomal translocations in human cells. We applied PETI to create cancer-associated translocations and inversions such as NPM1-ALK and EML4-ALK in human cells. Our findings show that PETI generated chromosomal translocation and inversion in a programmable manner with efficiencies comparable of Cas9. PETI methods, we believe, could be used to create disease models or for gene therapy.
科研通智能强力驱动
Strongly Powered by AbleSci AI