Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation

Abstract The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has facilitated dramatic progress in the field of genome engineering. Whilst microinjection of the Cas9 protein and a single guide RNA (sgRNA) into mouse zygotes is a widespread method for producing genetical...

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Autores principales:	Satoru Iwata, Hitomi Nakadai, Daisuke Fukushi, Mami Jose, Miki Nagahara, Takashi Iwamoto
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/253e25a047044934a4003b6259c44dd2
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spelling	oai:doaj.org-article:253e25a047044934a4003b6259c44dd22021-12-02T15:08:10ZSimple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation10.1038/s41598-019-50900-y2045-2322https://doaj.org/article/253e25a047044934a4003b6259c44dd22019-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-50900-yhttps://doaj.org/toc/2045-2322Abstract The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has facilitated dramatic progress in the field of genome engineering. Whilst microinjection of the Cas9 protein and a single guide RNA (sgRNA) into mouse zygotes is a widespread method for producing genetically engineered mice, in vitro and in vivo electroporation (which are much more convenient strategies) have recently been developed. However, it remains unknown whether these electroporation methods are able to manipulate genomes at the chromosome level. In the present study, we used these techniques to introduce chromosomal inversions of several megabases (Mb) in length in mouse zygotes. Using in vitro electroporation, we successfully introduced a 7.67 Mb inversion, which is longer than any previously reported inversion produced using microinjection-based methods. Additionally, using in vivo electroporation, we also introduced a long chromosomal inversion by targeting an allele in F1 hybrid mice. To our knowledge, the present study is the first report of target-specific chromosomal inversions in mammalian zygotes using electroporation.Satoru IwataHitomi NakadaiDaisuke FukushiMami JoseMiki NagaharaTakashi IwamotoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-8 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Satoru Iwata Hitomi Nakadai Daisuke Fukushi Mami Jose Miki Nagahara Takashi Iwamoto Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation
description	Abstract The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has facilitated dramatic progress in the field of genome engineering. Whilst microinjection of the Cas9 protein and a single guide RNA (sgRNA) into mouse zygotes is a widespread method for producing genetically engineered mice, in vitro and in vivo electroporation (which are much more convenient strategies) have recently been developed. However, it remains unknown whether these electroporation methods are able to manipulate genomes at the chromosome level. In the present study, we used these techniques to introduce chromosomal inversions of several megabases (Mb) in length in mouse zygotes. Using in vitro electroporation, we successfully introduced a 7.67 Mb inversion, which is longer than any previously reported inversion produced using microinjection-based methods. Additionally, using in vivo electroporation, we also introduced a long chromosomal inversion by targeting an allele in F1 hybrid mice. To our knowledge, the present study is the first report of target-specific chromosomal inversions in mammalian zygotes using electroporation.
format	article
author	Satoru Iwata Hitomi Nakadai Daisuke Fukushi Mami Jose Miki Nagahara Takashi Iwamoto
author_facet	Satoru Iwata Hitomi Nakadai Daisuke Fukushi Mami Jose Miki Nagahara Takashi Iwamoto
author_sort	Satoru Iwata
title	Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation
title_short	Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation
title_full	Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation
title_fullStr	Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation
title_full_unstemmed	Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation
title_sort	simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/253e25a047044934a4003b6259c44dd2
work_keys_str_mv	AT satoruiwata simpleandlargescalechromosomalengineeringofmousezygotesviainvitroandinvivoelectroporation AT hitominakadai simpleandlargescalechromosomalengineeringofmousezygotesviainvitroandinvivoelectroporation AT daisukefukushi simpleandlargescalechromosomalengineeringofmousezygotesviainvitroandinvivoelectroporation AT mamijose simpleandlargescalechromosomalengineeringofmousezygotesviainvitroandinvivoelectroporation AT mikinagahara simpleandlargescalechromosomalengineeringofmousezygotesviainvitroandinvivoelectroporation AT takashiiwamoto simpleandlargescalechromosomalengineeringofmousezygotesviainvitroandinvivoelectroporation
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Simple and large-scale chromosomal engineering of mouse zygotes via in vitro and in vivo electroporation

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