{"id":"doi:10.1186/s12896-016-0234-4","kind":"article","title":"Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes","authors":["Chu VT","Weber T","Graf R","Sommermann T","Petsch K","Sack U","Volchkov P","Rajewsky K","Kühn R"],"author_count":9,"year":2016,"published":"2016-01-16","venue":"BMC biotechnology","cited_by":281,"doi":"10.1186/s12896-016-0234-4","arxiv_id":null,"nct_id":null,"source_url":"https://doi.org/10.1186/s12896-016-0234-4","archive_url":"https://space-kitty.com/papers/doi:10.1186/s12896-016-0234-4","updated":null,"pmid":"26772810","pmcid":"PMC4715285","openalex_id":null,"pdf_url":"https://europepmc.org/articles/PMC4715285/pdf","abstract":"Background The CRISPR/Cas9 system is increasingly used for gene inactivation in mouse zygotes, but homology-directed mutagenesis and use of inbred embryos are less established. In particular, Rosa26 knock-in alleles for the insertion of transgenes in a genomic 'safe harbor' site, have not been produced. Here we applied CRISPR/Cas9 for the knock-in of 8-11 kb inserts into Rosa26 of C57BL/6 zygotes. Results We found that 10-20 % of live pups derived from microinjected zygotes were founder mutants, without apparent off-target effects, and up to 50 % knock-in embryos were recovered upon coinjection of Cas9 mRNA and protein. Using this approach, we established a new mouse line for the Cre/loxP-dependent expression of Cas9. Conclusions Altogether, our protocols and resources support the fast and direct generation of new Rosa26 knock-in alleles and of Cas9-mediated in vivo gene editing in the widely used C57BL/6 inbred strain.","abstract_license":"CC-BY (article license as recorded by Europe PMC; abstract reproduced unmodified apart from markup removal)","license":"CC-BY","is_open_access":true,"retracted":false,"peer_reviewed":null,"categories":[],"details":null,"provenance":"europepmc","attribution":"Abstract of \"Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes\" by Chu VT, Weber T, Graf R et al., https://doi.org/10.1186/s12896-016-0234-4, used under its Creative Commons Attribution license (CC-BY); markup removed, otherwise unmodified.","topics":[{"slug":"gene-editing-therapies","name":"CRISPR & Gene-Editing Therapies","domain_slug":"medicine"}],"cited_in_dossiers":[]}