Efficient design and assembly of custom TALENS using the golden gate platform

Tomas Cermak; Colby Starker; Daniel F Voytas

doi:10.1007/978-1-4939-1862-1_7

Efficient design and assembly of custom TALENS using the golden gate platform

Tomas Cermak, Colby Starker, Daniel F Voytas

Research output: Contribution to journal › Article › peer-review

Abstract

An important breakthrough in the field of genome engineering was the discovery of the modular T ranscription A ctivatorL ike E ffector (TALE) DNA binding domain and the development of TALE n ucleases (TALENs). TALENs enable researchers to make DNA double-strand breaks in target loci to create gene knockouts or introduce specific DNA sequence modifications. Precise genome engineering is increasingly being used to study gene function, develop disease models or create new traits in crop species. Underlying the boom in genome engineering is the striking simplicity and low cost of engineering new specificities of TALENs and other sequence-specific nucleases. In this chapter, we describe a rapid, inexpensive, and user-friendly protocol for custom TALEN construction based on one of the most popular TALEN assembly platforms, the Golden Gate cloning method. Using this protocol, ready-to-use TALENs with specificity for targets 13-32 bp long are constructed within 5 days.

Original language	English (US)
Pages (from-to)	133-159
Number of pages	27
Journal	Methods in Molecular Biology
Volume	1239
DOIs	https://doi.org/10.1007/978-1-4939-1862-1_7
State	Published - Jan 1 2015

Keywords

Argeted mutagenesis
Genome engineering
Golden Gate assembly
Site-specific nuclease
TALEN

Access

10.1007/978-1-4939-1862-1_7

OpenUrl availability

Full text

Cite this

@article{b06e396f04cc4a0590c389565c852033,

title = "Efficient design and assembly of custom TALENS using the golden gate platform",

abstract = "An important breakthrough in the field of genome engineering was the discovery of the modular T ranscription A ctivatorL ike E ffector (TALE) DNA binding domain and the development of TALE n ucleases (TALENs). TALENs enable researchers to make DNA double-strand breaks in target loci to create gene knockouts or introduce specific DNA sequence modifications. Precise genome engineering is increasingly being used to study gene function, develop disease models or create new traits in crop species. Underlying the boom in genome engineering is the striking simplicity and low cost of engineering new specificities of TALENs and other sequence-specific nucleases. In this chapter, we describe a rapid, inexpensive, and user-friendly protocol for custom TALEN construction based on one of the most popular TALEN assembly platforms, the Golden Gate cloning method. Using this protocol, ready-to-use TALENs with specificity for targets 13-32 bp long are constructed within 5 days.",

keywords = "Argeted mutagenesis, Genome engineering, Golden Gate assembly, Site-specific nuclease, TALEN",

author = "Tomas Cermak and Colby Starker and Voytas, {Daniel F}",

year = "2015",

month = jan,

day = "1",

doi = "10.1007/978-1-4939-1862-1_7",

language = "English (US)",

volume = "1239",

pages = "133--159",

journal = "Methods in Molecular Biology",

issn = "1064-3745",

publisher = "Humana Press",

}

TY - JOUR

T1 - Efficient design and assembly of custom TALENS using the golden gate platform

AU - Cermak, Tomas

AU - Starker, Colby

AU - Voytas, Daniel F

PY - 2015/1/1

Y1 - 2015/1/1

N2 - An important breakthrough in the field of genome engineering was the discovery of the modular T ranscription A ctivatorL ike E ffector (TALE) DNA binding domain and the development of TALE n ucleases (TALENs). TALENs enable researchers to make DNA double-strand breaks in target loci to create gene knockouts or introduce specific DNA sequence modifications. Precise genome engineering is increasingly being used to study gene function, develop disease models or create new traits in crop species. Underlying the boom in genome engineering is the striking simplicity and low cost of engineering new specificities of TALENs and other sequence-specific nucleases. In this chapter, we describe a rapid, inexpensive, and user-friendly protocol for custom TALEN construction based on one of the most popular TALEN assembly platforms, the Golden Gate cloning method. Using this protocol, ready-to-use TALENs with specificity for targets 13-32 bp long are constructed within 5 days.

AB - An important breakthrough in the field of genome engineering was the discovery of the modular T ranscription A ctivatorL ike E ffector (TALE) DNA binding domain and the development of TALE n ucleases (TALENs). TALENs enable researchers to make DNA double-strand breaks in target loci to create gene knockouts or introduce specific DNA sequence modifications. Precise genome engineering is increasingly being used to study gene function, develop disease models or create new traits in crop species. Underlying the boom in genome engineering is the striking simplicity and low cost of engineering new specificities of TALENs and other sequence-specific nucleases. In this chapter, we describe a rapid, inexpensive, and user-friendly protocol for custom TALEN construction based on one of the most popular TALEN assembly platforms, the Golden Gate cloning method. Using this protocol, ready-to-use TALENs with specificity for targets 13-32 bp long are constructed within 5 days.

KW - Argeted mutagenesis

KW - Genome engineering

KW - Golden Gate assembly

KW - Site-specific nuclease

KW - TALEN

UR - http://www.scopus.com/inward/record.url?scp=84911942784&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84911942784&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-1862-1_7

DO - 10.1007/978-1-4939-1862-1_7

M3 - Article

C2 - 25408404

SN - 1064-3745

VL - 1239

SP - 133

EP - 159

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

Efficient design and assembly of custom TALENS using the golden gate platform

Abstract

Keywords

Access

OpenUrl availability

Other files and links

Fingerprint

Cite this