Generation of eGFP and Cre knockin rats by CRISPR/Cas9.

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  • Additional Information
    • Source:
      Publisher: Published by Blackwell Pub. on behalf of the Federation of European Biochemical Societies Country of Publication: England NLM ID: 101229646 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1742-4658 (Electronic) Linking ISSN: 1742464X NLM ISO Abbreviation: FEBS J Subsets: MEDLINE
    • Publication Information:
      Original Publication: Oxford, UK : Published by Blackwell Pub. on behalf of the Federation of European Biochemical Societies, c2005-
    • Subject Terms:
      Clustered Regularly Interspaced Short Palindromic Repeats*; Gene Knock-In Techniques/*methods ; Green Fluorescent Proteins/*genetics ; Integrases/*genetics; Animals ; Base Sequence ; Genetic Engineering/methods ; Genetic Vectors ; Germ Cells ; Nestin/genetics ; Rats
    • Abstract:
      The type II bacterial CRISPR/Cas [clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas)] system is a very valuable genome engineering tool, which has been widely used in genome editing of a variety of organisms. Previously, we generated floxed alleles in rats by CRISPR/Cas9. Here, we successfully use a two-cut strategy with one circular vector, which contains the exogenous cDNAs with homology arm regions, in generating knockin rats at the Trdmt1, Nestin and Cck loci. The efficiency of CRISPR/Cas9-mediated knockin was up to 54%. Furthermore, by crossing the Nestin-Cre rat with the Dnmt3b floxed rat and Cck-Cre with the Dnmt1 floxed rat, we detected Cre/loxP-mediated recombination in the F1 generation of rats. We also show that the knockin alleles were germline transmitted. These results provided a simple and flexible engineering strategy for the establishment of knockin rats.
      (© 2014 FEBS.)
    • Contributed Indexing:
      Keywords: CRISPR/Cas9; Cck; knockin; nestin, rat; trdmt1
    • Accession Number:
      0 (Nes protein, rat)
      0 (Nestin)
      0 (enhanced green fluorescent protein)
      147336-22-9 (Green Fluorescent Proteins)
      EC 2.7.7.- (Cre recombinase)
      EC 2.7.7.- (Integrases)
    • Publication Date:
      Date Created: 20140722 Date Completed: 20141104 Latest Revision: 20140904
    • Publication Date:
      20240829
    • Accession Number:
      10.1111/febs.12935
    • Accession Number:
      25039742