Genome engineering with custom recombinases

Thomas Gaj, Carlos F. Barbas

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Site-specific recombinases are valuable tools for myriad basic research and genome engineering applications. In particular, hybrid recombinases consisting of catalytic domains from the resolvase/invertase family of serine recombinases fused to Cys2-His2 zinc-finger or TAL effector DNA-binding domains are capable of introducing targeted modifications into mammalian cells. Due to their inherent modularity, new recombinases with distinct targeting specificities can readily be generated and utilized in a "plug-and-play" manner. In this protocol, we provide detailed, step-by-step instructions for generating new hybrid recombinases with user-defined specificity, as well as methods for achieving site-specific integration into targeted genomic loci using these systems.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages79-91
Number of pages13
EditionC
DOIs
StatePublished - 2014

Publication series

NameMethods in Enzymology
NumberC
Volume546
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Genome engineering
  • Protein engineering
  • Recombinase
  • TAL effector
  • Zinc-finger

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Gaj, T., & Barbas, C. F. (2014). Genome engineering with custom recombinases. In Methods in Enzymology (C ed., pp. 79-91). (Methods in Enzymology; Vol. 546, No. C). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-801185-0.00004-0