Successful <i>i</i>-GONAD in Mice at Early Zygote Stage through In Vivo Electroporation Three Min after Intraoviductal Instillation of CRISPR-Ribonucleoprotein by Shuji Takabayashi, Kenta Iijima, Masumi Tsujimura, Takuya Aoshima, Hisayoshi Takagi, Kazushi Aoto, Masahiro Sato

Subjects: Get full text

Recent Advances and Future Perspectives of In Vivo Targeted Delivery of Genome-Editing Reagents to Germ Cells, Embryos, and Fetuses in Mice by Masahiro Sato, Shuji Takabayashi, Eri Akasaka, Shingo Nakamura

Subjects: Get full text

Recent Advances in the Production of Genome-Edited Rats by Masahiro Sato, Shingo Nakamura, Emi Inada, Shuji Takabayashi

“…In vitro electroporation (EP) of zygotes is next recognized as a simple and rapid method to introduce GE components to produce GE animals. …”
Get full text

Sequential i-GONAD: An Improved In Vivo Technique for CRISPR/Cas9-Based Genetic Manipulations in Mice by Masahiro Sato, Rico Miyagasako, Shuji Takabayashi, Masato Ohtsuka, Izuho Hatada, Takuro Horii

“…Improved genome-editing via oviductal nucleic acid delivery (i-GONAD) is a technique capable of inducing genomic changes in preimplantation embryos (zygotes) present within the oviduct of a pregnant female. i-GONAD involves intraoviductal injection of a solution containing genome-editing components via a glass micropipette under a dissecting microscope, followed by in vivo electroporation using tweezer-type electrodes. i-GONAD does not involve ex vivo handling of embryos (isolation of zygotes, microinjection or electroporation of zygotes, and egg transfer of the treated embryos to the oviducts of a recipient female), which is required for in vitro genome-editing of zygotes. i-GONAD enables the generation of indels, knock-in (KI) of ~ 1 kb sequence of interest, and large deletion at a target locus. i-GONAD is usually performed on Day 0.7 of pregnancy, which corresponds to the late zygote stage. …”
Get full text

Modification of <i>i</i>-GONAD Suitable for Production of Genome-Edited C57BL/6 Inbred Mouse Strain by Yukari Kobayashi, Takuya Aoshima, Ryota Ito, Ryota Shinmura, Masato Ohtsuka, Eri Akasaka, Masahiro Sato, Shuji Takabayashi

“…Improved genome editing via oviductal nucleic acid delivery (<i>i</i>-GONAD) is a novel method for producing genome-edited mice in the absence of ex vivo handling of zygotes. <i>i</i>-GONAD involves the intraoviductal injection of clustered regularly interspaced short palindromic repeats (CRISPR) ribonucleoproteins via the oviductal wall of pregnant females at 0.7 days post-coitum, followed by in vivo electroporation (EP). …”
Get full text