@article{d269e280d81b4dc4bb0ea2080006cf91,
title = "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.",
keywords = "CRISPR, Easi-CRISPR, GONAD, In vivo electroporation, Knock-in, Long ssDNA, Transgenic mouse",
author = "Masato Ohtsuka and Masahiro Sato and Hiromi Miura and Shuji Takabayashi and Makoto Matsuyama and Takayuki Koyano and Naomi Arifin and Shingo Nakamura and Kenta Wada and Gurumurthy, {Channabasavaiah B.}",
note = "Funding Information: This work was supported in part by a Grant-in-Aid for Young Scientists (B) (16K18821) from the Japan Society for the Promotion of Science (JSPS) to H.M. M.O. acknowledges the funding support from the 2014 Tokai University School of Medicine Research Aid, MEXT-Supported Program for the Strategic Research Foundation at Private Universities 2015–2019 (PI: Y.Inagaki), Research and Study Project of Tokai University General Research Organization, 2016–2017 Tokai University School of Medicine Project Research, and a Grant-in-Aid for challenging Exploratory Research (15K14371) from JSPS. This work was also supported by a Grant-in-Aid for Scientific Research (B) (16H05049) from JSPS to S.N. Publisher Copyright: {\textcopyright} 2018 The Author(s).",
year = "2018",
month = feb,
day = "26",
doi = "10.1186/s13059-018-1400-x",
language = "English (US)",
volume = "19",
journal = "Genome biology",
issn = "1474-7596",
publisher = "BioMed Central",
number = "1",
}