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

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.

Original language	English
Pages (from-to)	853-9
Number of pages	7
Journal	ACS Synthetic Biology
Volume	4
Issue number	7
DOIs	https://doi.org/10.1021/sb500372z
Publication status	Published - 17 Jul 2015

Keywords

3' Untranslated Regions
DNA
Genetic Engineering
Genetic Vectors
Open Reading Frames
Promoter Regions, Genetic
Saccharomyces cerevisiae
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

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10.1021/sb500372z

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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.",

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

year = "2015",

month = jul,

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doi = "10.1021/sb500372z",

language = "English",

volume = "4",

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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.

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.

KW - 3' Untranslated Regions

KW - DNA

KW - Genetic Engineering

KW - Genetic Vectors

KW - Open Reading Frames

KW - Promoter Regions, Genetic

KW - Saccharomyces cerevisiae

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

KW - Research Support, U.S. Gov't, Non-P.H.S.

U2 - 10.1021/sb500372z

DO - 10.1021/sb500372z

M3 - Article

C2 - 25756291

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

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