Gene-targeting vectors

J. Simon, C. Arthur, Victoria A. McGuire

    Research output: Chapter in Book/Report/Conference proceedingChapter

    4 Citations (Scopus)

    Abstract

    Gene targeting in mice has been used extensively to elucidate gene function in vivo. However, for gene targeting to be successful, the targeting vector must be carefully designed. This chapter addresses the rationale behind designing targeting vectors, detailing the essential components, and highlighting specific considerations for different types of vectors, from gene deletions to point mutations and insertions. Examples of vector designs, cloning strategies, and approaches for successful screening of recombinants are described. The use of Cre/LoxP and Flp/frt systems for conditional targeting is described, together with strategies for generating conditional deletions. Methods for generating conditional point mutations are also described and their potential drawbacks discussed.

    Original languageEnglish
    Title of host publicationTransgenesis techniques
    Subtitle of host publicationprinciples and protocols
    EditorsElizabeth J. Cartwright
    PublisherSpringer
    Pages127-144
    Number of pages18
    Edition3
    ISBN (Electronic)978-1-60327-019-9
    ISBN (Print)9781603270182
    DOIs
    Publication statusPublished - May 2009

    Publication series

    NameMethods in Molecular Biology
    Volume561
    ISSN (Print)1064-3745

    Keywords

    • Cre
    • Frt
    • Gene-targeting vector
    • Knockin
    • Knockout

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

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