   3.5.4.38 | molecular biology |
a target-AID base editor, designed to recruit cytidine deaminase (CDA) to the target DNA locus via the CRISPR/Cas9 system, can directly induce C to T mutation without double-strand breaks and donor DNA. This system is adopted in Yarrowia lipolytica for multiplex gene disruption. Target-specific gRNA(s) and a fusion protein consisting of a nickase Cas9, CDA1, and uracil DNA glycosylase inhibitor are expressed from a single plasmid to disrupt target genes by introducing a stop codon via C to T mutation within the mutational window. Using this Target-AID system, single gene disruption and simultaneous double gene disruption are achieved with the efficiencies up to 94% and 31%, respectively |
753049 |
