ErCas12a CRISPR-MAD7 for Model Generation in Human Cells, Mice, and Rats
Zhenyi Liu, John A. Schiel, Elena Maksimova, Žaklina Strezoska, Guojun Zhao, Emily M. Anderson, Yumei Wu, Joe Warren, Angela Bartels, Anja van Brabant Smith, Chris Lowe, Kevin P. Forbes
Abstract
MAD7 is an engineered class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) system isolated from Eubacterium rectale. Analogous to Cas9, it is an RNA-guided nuclease with demonstrated gene editing activity in Escherichia coli and yeast cells. Here, we report that MAD7 is capable of generating indels and fluorescent gene tagging of endogenous genes in human HCT116 and U2OS cancer cell lines, respectively. In addition, MAD7 is highly proficient in generating indels, small DNA insertions (23 bases), and larger integrations ranging from 1 to 14 kb in size in mouse and rat embryos, resulting in live-born transgenic animals. Due to the different protospacer adjacent motif requirement, small-guide RNA, and highly efficient targeted gene disruption and insertions, MAD7 can expand the CRISPR toolbox for genome enginnering across different systems and model organisms.