Mouse Genome Editing from Scratch

Abstract

Mouse genome modification requires costly equipment and highly skilled personnel to manipulate zygotes. A number of zygote electroporation techniques were reported to be highly efficient in gene delivery. One of these methods called i-GONAD (improved Genome-editing via Oviductal Nucleic Acids Delivery) describes electroporation-based gene transfer to zygotes in utero. Here we adopted this technology for mouse genome-editing from scratch minimizing the cost of equipment, operator skill and animal use. We chose the CRISPR/Cas9 system as a genome editing tool and i-GONAD as a gene delivery method to produce IL10 gene knockout in C57BL/6 mice. Four animals out of 13 delivered pups (30.8%) were genetically compromised at IL10 gene locus suggesting the feasibility of the approach. This report provides one of the possible technical settings for those who aim at establishing in-house mouse transgenesis pipeline at minimal cost from scratch. For citationPopova J.V., Bets V.D., Omelina E.S., Boldyreva L.V., Kozhevnikova E.N. Mouse Genome Editing from Scratch. Journal of Experimental Biology. Year; issue (number): page. DOI