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

Fingerprint

Recombinases
Genome
Catalytic Domain
DNA
Zinc Fingers
Research
Serine
Research Personnel
Enzymes

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

Cite this

Sirk, S. (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

Directed evolution of targeted recombinases for genome engineering. / Sirk, Shannon.

Methods in Molecular Biology. Humana Press Inc., 2018. p. 89-102 (Methods in Molecular Biology; Vol. 1867).

Research output: Chapter in Book/Report/Conference proceedingChapter

Sirk, S 2018, Directed evolution of targeted recombinases for genome engineering. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1867, Humana Press Inc., pp. 89-102. https://doi.org/10.1007/978-1-4939-8799-3_7
Sirk S. Directed evolution of targeted recombinases for genome engineering. In Methods in Molecular Biology. Humana Press Inc. 2018. p. 89-102. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8799-3_7
Sirk, Shannon. / Directed evolution of targeted recombinases for genome engineering. Methods in Molecular Biology. Humana Press Inc., 2018. pp. 89-102 (Methods in Molecular Biology).
@inbook{7b4e2fbfe4644e85833727c7f5f75c13,
title = "Directed evolution of targeted recombinases for genome engineering",
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.",
keywords = "Directed evolution, Genetic engineering, Genome editing, Protein engineering, Rational design, Recombinase, Site-specific genomic modification, Targeted integration",
author = "Shannon Sirk",
year = "2018",
month = "1",
day = "1",
doi = "10.1007/978-1-4939-8799-3_7",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "89--102",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Directed evolution of targeted recombinases for genome engineering

AU - Sirk, Shannon

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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.

AB - 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.

KW - Directed evolution

KW - Genetic engineering

KW - Genome editing

KW - Protein engineering

KW - Rational design

KW - Recombinase

KW - Site-specific genomic modification

KW - Targeted integration

UR - http://www.scopus.com/inward/record.url?scp=85052717388&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85052717388&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-8799-3_7

DO - 10.1007/978-1-4939-8799-3_7

M3 - Chapter

C2 - 30155817

AN - SCOPUS:85052717388

T3 - Methods in Molecular Biology

SP - 89

EP - 102

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -