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

M Ohtsuka, M Sato, H Miura, S Takabayashi… - Genome biology, 2018 - Springer
M Ohtsuka, M Sato, H Miura, S Takabayashi, M Matsuyama, T Koyano, N Arifin, S Nakamura…
Genome biology, 2018Springer
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, which rely on ex vivo handling of zygotes and require
recipient animals for embryo transfer. In contrast, i-GONAD avoids these technically difficult …
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, 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, i-GONAD-treated females retain reproductive function, suggesting future use of the method for germline gene therapy.
Springer