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


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.

SUBMITTER: Ohtsuka M 

PROVIDER: S-EPMC5828090 | biostudies-other | 2018 Feb

REPOSITORIES: biostudies-other

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

Ohtsuka Masato M   Sato Masahiro M   Miura Hiromi H   Takabayashi Shuji S   Matsuyama Makoto M   Koyano Takayuki T   Arifin Naomi N   Nakamura Shingo S   Wada Kenta K   Gurumurthy Channabasavaiah B CB  

Genome biology 20180226 1


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 av  ...[more]

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