Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units

Neta Agmon; Leslie A. Mitchell; Yizhi Cai; Shigehito Ikushima; James Chuang; Allen Zheng; Woo Jin Choi; J. Andrew Martin; Katrina Caravelli; Giovanni Stracquadanio; Jef D. Boeke

doi:10.1021/sb500372z

Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units

Neta Agmon, Leslie A. Mitchell, Yizhi Cai, Shigehito Ikushima, James Chuang, Allen Zheng, Woo Jin Choi, J. Andrew Martin, Katrina Caravelli, Giovanni Stracquadanio, Jef D. Boeke

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

49 Scopus citations

Abstract

We have adapted the Golden Gate DNA assembly method to the assembly of transcription units (TUs) for the yeast Saccharomyces cerevisiae, in a method we call yeast Golden Gate (yGG). yGG allows for the easy assembly of TUs consisting of promoters (PRO), coding sequences (CDS), and terminators (TER). Carefully designed overhangs exposed by digestion with a type IIS restriction enzyme enable virtually seamless assembly of TUs that, in principle, contain all of the information necessary to express a gene of interest in yeast. We also describe a versatile set of yGG acceptor vectors to be used for TU assembly. These vectors can be used for low or high copy expression of assembled TUs or integration into carefully selected innocuous genomic loci. yGG provides synthetic biologists and yeast geneticists with an efficient new means by which to engineer S. cerevisiae. (Figure Presented).

Original language	English (US)
Pages (from-to)	853-859
Number of pages	7
Journal	ACS Synthetic Biology
Volume	4
Issue number	7
DOIs	https://doi.org/10.1021/sb500372z
State	Published - Jul 17 2015
Externally published	Yes

Keywords

DNA assembly
S. cerevisiae
synthetic biology
transcription unit
yeast Golden Gate

ASJC Scopus subject areas

Biomedical Engineering
Biochemistry, Genetics and Molecular Biology (miscellaneous)

Access to Document

10.1021/sb500372z

Cite this

@article{9275209cbc14463688cf4dd52c3fc6a5,

title = "Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units",

abstract = "We have adapted the Golden Gate DNA assembly method to the assembly of transcription units (TUs) for the yeast Saccharomyces cerevisiae, in a method we call yeast Golden Gate (yGG). yGG allows for the easy assembly of TUs consisting of promoters (PRO), coding sequences (CDS), and terminators (TER). Carefully designed overhangs exposed by digestion with a type IIS restriction enzyme enable virtually seamless assembly of TUs that, in principle, contain all of the information necessary to express a gene of interest in yeast. We also describe a versatile set of yGG acceptor vectors to be used for TU assembly. These vectors can be used for low or high copy expression of assembled TUs or integration into carefully selected innocuous genomic loci. yGG provides synthetic biologists and yeast geneticists with an efficient new means by which to engineer S. cerevisiae. (Figure Presented).",

keywords = "DNA assembly, S. cerevisiae, synthetic biology, transcription unit, yeast Golden Gate",

author = "Neta Agmon and Mitchell, {Leslie A.} and Yizhi Cai and Shigehito Ikushima and James Chuang and Allen Zheng and Choi, {Woo Jin} and Martin, {J. Andrew} and Katrina Caravelli and Giovanni Stracquadanio and Boeke, {Jef D.}",

note = "Publisher Copyright: {\textcopyright} 2015 American Chemical Society.",

year = "2015",

month = jul,

day = "17",

doi = "10.1021/sb500372z",

language = "English (US)",

volume = "4",

pages = "853--859",

journal = "ACS Synthetic Biology",

issn = "2161-5063",

publisher = "American Chemical Society",

number = "7",

}

TY - JOUR

T1 - Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units

AU - Agmon, Neta

AU - Mitchell, Leslie A.

AU - Cai, Yizhi

AU - Ikushima, Shigehito

AU - Chuang, James

AU - Zheng, Allen

AU - Choi, Woo Jin

AU - Martin, J. Andrew

AU - Caravelli, Katrina

AU - Stracquadanio, Giovanni

AU - Boeke, Jef D.

PY - 2015/7/17

Y1 - 2015/7/17

N2 - We have adapted the Golden Gate DNA assembly method to the assembly of transcription units (TUs) for the yeast Saccharomyces cerevisiae, in a method we call yeast Golden Gate (yGG). yGG allows for the easy assembly of TUs consisting of promoters (PRO), coding sequences (CDS), and terminators (TER). Carefully designed overhangs exposed by digestion with a type IIS restriction enzyme enable virtually seamless assembly of TUs that, in principle, contain all of the information necessary to express a gene of interest in yeast. We also describe a versatile set of yGG acceptor vectors to be used for TU assembly. These vectors can be used for low or high copy expression of assembled TUs or integration into carefully selected innocuous genomic loci. yGG provides synthetic biologists and yeast geneticists with an efficient new means by which to engineer S. cerevisiae. (Figure Presented).

AB - We have adapted the Golden Gate DNA assembly method to the assembly of transcription units (TUs) for the yeast Saccharomyces cerevisiae, in a method we call yeast Golden Gate (yGG). yGG allows for the easy assembly of TUs consisting of promoters (PRO), coding sequences (CDS), and terminators (TER). Carefully designed overhangs exposed by digestion with a type IIS restriction enzyme enable virtually seamless assembly of TUs that, in principle, contain all of the information necessary to express a gene of interest in yeast. We also describe a versatile set of yGG acceptor vectors to be used for TU assembly. These vectors can be used for low or high copy expression of assembled TUs or integration into carefully selected innocuous genomic loci. yGG provides synthetic biologists and yeast geneticists with an efficient new means by which to engineer S. cerevisiae. (Figure Presented).

KW - DNA assembly

KW - S. cerevisiae

KW - synthetic biology

KW - transcription unit

KW - yeast Golden Gate

UR - http://www.scopus.com/inward/record.url?scp=84937157618&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84937157618&partnerID=8YFLogxK

U2 - 10.1021/sb500372z

DO - 10.1021/sb500372z

M3 - Article

C2 - 25756291

AN - SCOPUS:84937157618

SN - 2161-5063

VL - 4

SP - 853

EP - 859

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

IS - 7

ER -

Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this