Directed evolution of targeted recombinases for genome engineering

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Over the past several years, genome engineering has become an established component of basic research endeavors, and is emerging as a vital element of clinical research applications. Site-specific recombinases are one of the several tools that can facilitate genome modification by catalyzing rearrangements between specific DNA targets. Of particular interest are the small serine recombinases, which are modular in both form and function. This unique structure permits replacement of the native DNA-binding domain with designer targeting modules such as zinc fingers, TALEs, or catalytically inactivated Cas9, enabling modification of investigator-defined genomic loci. Importantly, the catalytic domain of these enzymes also contributes to target specificity, and can be reprogrammed to recognize custom sequences for genomic targeting. Here we describe the steps required to construct, select, and validate hybrid recombinase catalytic domains for targeted genome engineering.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages89-102
Number of pages14
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1867
ISSN (Print)1064-3745

Keywords

  • Directed evolution
  • Genetic engineering
  • Genome editing
  • Protein engineering
  • Rational design
  • Recombinase
  • Site-specific genomic modification
  • Targeted integration

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Directed evolution of targeted recombinases for genome engineering'. Together they form a unique fingerprint.

  • Cite this

    Sirk, S. J. (2018). Directed evolution of targeted recombinases for genome engineering. In Methods in Molecular Biology (pp. 89-102). (Methods in Molecular Biology; Vol. 1867). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8799-3_7