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...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2018-02-01
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| Series: | Genome Biology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13059-018-1400-x |
| _version_ | 1818311942433931264 |
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| author | Masato Ohtsuka Masahiro Sato Hiromi Miura Shuji Takabayashi Makoto Matsuyama Takayuki Koyano Naomi Arifin Shingo Nakamura Kenta Wada Channabasavaiah B. Gurumurthy |
| author_facet | Masato Ohtsuka Masahiro Sato Hiromi Miura Shuji Takabayashi Makoto Matsuyama Takayuki Koyano Naomi Arifin Shingo Nakamura Kenta Wada Channabasavaiah B. Gurumurthy |
| author_sort | Masato Ohtsuka |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T08:09:58Z |
| format | Article |
| id | doaj.art-434525404bbe47ed92639ed52ed7de0a |
| institution | Directory Open Access Journal |
| issn | 1474-760X |
| language | English |
| last_indexed | 2024-12-13T08:09:58Z |
| publishDate | 2018-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Genome Biology |
| spelling | doaj.art-434525404bbe47ed92639ed52ed7de0a2022-12-21T23:54:14ZengBMCGenome Biology1474-760X2018-02-0119111510.1186/s13059-018-1400-xi-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleasesMasato Ohtsuka0Masahiro Sato1Hiromi Miura2Shuji Takabayashi3Makoto Matsuyama4Takayuki Koyano5Naomi Arifin6Shingo Nakamura7Kenta Wada8Channabasavaiah B. Gurumurthy9Department of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, School of Medicine, Tokai UniversitySection of Gene Expression Regulation, Frontier Science Research Center, Kagoshima UniversityDepartment of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, School of Medicine, Tokai UniversityLaboratory Animal Facilities & Services, Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of MedicineDivision of Molecular Genetics, Shigei Medical Research InstituteDivision of Molecular Genetics, Shigei Medical Research InstituteDepartment of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, School of Medicine, Tokai UniversityDivision of Biomedical Engineering, National Defense Medical College Research InstituteDepartment of Bioproduction, Tokyo University of AgricultureMouse Genome Engineering Core Facility, Vice Chancellor for Research Office, University of Nebraska Medical CenterAbstract 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.http://link.springer.com/article/10.1186/s13059-018-1400-xIn vivo electroporationCRISPRGONADKnock-inTransgenic mouseLong ssDNA |
| spellingShingle | Masato Ohtsuka Masahiro Sato Hiromi Miura Shuji Takabayashi Makoto Matsuyama Takayuki Koyano Naomi Arifin Shingo Nakamura Kenta Wada Channabasavaiah B. Gurumurthy i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases Genome Biology In vivo electroporation CRISPR GONAD Knock-in Transgenic mouse Long ssDNA |
| title | i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases |
| title_full | i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases |
| title_fullStr | i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases |
| title_full_unstemmed | i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases |
| title_short | i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases |
| title_sort | i gonad a robust method for in situ germline genome engineering using crispr nucleases |
| topic | In vivo electroporation CRISPR GONAD Knock-in Transgenic mouse Long ssDNA |
| url | http://link.springer.com/article/10.1186/s13059-018-1400-x |
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