Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox
Summary: Precise integration of DNA constructs greater than 3 kb into mouse zygotes is difficult. Here, we present a protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox. We describe steps for choosing mouse strains with preplaced attachment sites. We then detail procedures for mi...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724001874 |
| _version_ | 1827281383840546816 |
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| author | Valerie Erhardt Chrystal M. Snow Vishnu Hosur |
| author_facet | Valerie Erhardt Chrystal M. Snow Vishnu Hosur |
| author_sort | Valerie Erhardt |
| collection | DOAJ |
| description | Summary: Precise integration of DNA constructs greater than 3 kb into mouse zygotes is difficult. Here, we present a protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox. We describe steps for choosing mouse strains with preplaced attachment sites. We then detail procedures for microinjecting mouse zygotes with the plasmid donor DNA construct to generate transgenic mice by recombination-mediated cassette exchange. This protocol has the potential for application in exploring the functional implications of large structural variations in cancer.For complete details on the use and execution of this protocol, please refer to Low et al.1 and Hosur et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-24T08:59:30Z |
| format | Article |
| id | doaj.art-40be595ab342459aab90c3b92b524383 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-24T08:59:30Z |
| publishDate | 2024-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-40be595ab342459aab90c3b92b5243832024-04-16T04:09:51ZengElsevierSTAR Protocols2666-16672024-06-0152103022Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolboxValerie Erhardt0Chrystal M. Snow1Vishnu Hosur2The Jackson Laboratory for Mammalian Genetics, Bar Harbor, ME, USAThe Jackson Laboratory for Mammalian Genetics, Bar Harbor, ME, USAThe Jackson Laboratory for Mammalian Genetics, Bar Harbor, ME, USA; Corresponding authorSummary: Precise integration of DNA constructs greater than 3 kb into mouse zygotes is difficult. Here, we present a protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox. We describe steps for choosing mouse strains with preplaced attachment sites. We then detail procedures for microinjecting mouse zygotes with the plasmid donor DNA construct to generate transgenic mice by recombination-mediated cassette exchange. This protocol has the potential for application in exploring the functional implications of large structural variations in cancer.For complete details on the use and execution of this protocol, please refer to Low et al.1 and Hosur et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724001874GeneticsCRISPRBiotechnology and bioengineering |
| spellingShingle | Valerie Erhardt Chrystal M. Snow Vishnu Hosur Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox STAR Protocols Genetics CRISPR Biotechnology and bioengineering |
| title | Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox |
| title_full | Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox |
| title_fullStr | Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox |
| title_full_unstemmed | Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox |
| title_short | Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox |
| title_sort | protocol for large dna transgenesis in mice using the cas9 bxb1 toolbox |
| topic | Genetics CRISPR Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724001874 |
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