i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases

i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases

Blog Article

Abstract We present a robust method called improved-Genome editing via Oviductal Nucleic Acids Delivery (i-GONAD) that delivers CRISPR ribonucleoproteins to E0.7 embryos via in situ electroporation.The method generates mouse models containing single-base changes, kilobase-sized deletions, and knock-ins.The efficiency of i-GONAD is comparable to that of traditional microinjection methods, jmannino.com which rely on ex vivo handling of zygotes and require recipient animals for embryo transfer.

In contrast, i-GONAD avoids these technically difficult steps, and it can be performed at any laboratory with simple equipment and technical expertise.Further, here i-GONAD-treated females retain reproductive function, suggesting future use of the method for germline gene therapy.