Multiple-gene silencing using antisense RNAs in Escherichia coli

Nobutaka Nakashima; Shan Goh; Liam Good; Tomohiro Tamura

doi:10.1007/978-1-61779-424-7_23

Multiple-gene silencing using antisense RNAs in Escherichia coli

Nobutaka Nakashima, Shan Goh, Liam Good, Tomohiro Tamura

Research output: Contribution to journal › Article

15 Citations (Scopus)

Abstract

We have developed four expression vectors to express antisense RNAs (asRNAs) by which genes of interest are silenced in Escherichia coli. The vectors are all IPTG-inducible and co-transformable in any combination and target genes are silenced conditionally and concurrently. Furthermore, in order to improve silencing efficacy, the vectors are designed to express uniquely shaped antisense RNAs, named paired termini antisense RNAs (PTasRNAs). The vectors are useful for comprehensive investigation of gene function and are applicable even if the target genes are essential for cell growth. Here, we describe methods to construct PTasRNA-expressing vectors and to evaluate silencing efficacy.

Original language	English
Pages (from-to)	307-19
Number of pages	13
Journal	Methods in Molecular Biology
Volume	815
DOIs	https://doi.org/10.1007/978-1-61779-424-7_23
Publication status	Published - 2012

Keywords

Culture Techniques
DNA, Bacterial
DNA, Recombinant
Escherichia coli
Escherichia coli Proteins
Gene Expression Regulation, Bacterial
Gene Knockdown Techniques
Gene Silencing
Genetic Engineering
Inverted Repeat Sequences
Molecular Biology
Molecular Sequence Data
Polymerase Chain Reaction
RNA, Antisense
Transformation, Bacterial
Journal Article
Research Support, Non-U.S. Gov't

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10.1007/978-1-61779-424-7_23

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title = "Multiple-gene silencing using antisense RNAs in Escherichia coli",

abstract = "We have developed four expression vectors to express antisense RNAs (asRNAs) by which genes of interest are silenced in Escherichia coli. The vectors are all IPTG-inducible and co-transformable in any combination and target genes are silenced conditionally and concurrently. Furthermore, in order to improve silencing efficacy, the vectors are designed to express uniquely shaped antisense RNAs, named paired termini antisense RNAs (PTasRNAs). The vectors are useful for comprehensive investigation of gene function and are applicable even if the target genes are essential for cell growth. Here, we describe methods to construct PTasRNA-expressing vectors and to evaluate silencing efficacy.",

keywords = "Culture Techniques, DNA, Bacterial, DNA, Recombinant, Escherichia coli, Escherichia coli Proteins, Gene Expression Regulation, Bacterial, Gene Knockdown Techniques, Gene Silencing, Genetic Engineering, Inverted Repeat Sequences, Molecular Biology, Molecular Sequence Data, Polymerase Chain Reaction, RNA, Antisense, Transformation, Bacterial, Journal Article, Research Support, Non-U.S. Gov't",

author = "Nobutaka Nakashima and Shan Goh and Liam Good and Tomohiro Tamura",

year = "2012",

doi = "10.1007/978-1-61779-424-7_23",

language = "English",

volume = "815",

pages = "307--19",

journal = "Methods in Molecular Biology",

issn = "1064-3745",

publisher = "Humana Press",

}

TY - JOUR

T1 - Multiple-gene silencing using antisense RNAs in Escherichia coli

AU - Nakashima, Nobutaka

AU - Goh, Shan

AU - Good, Liam

AU - Tamura, Tomohiro

PY - 2012

Y1 - 2012

N2 - We have developed four expression vectors to express antisense RNAs (asRNAs) by which genes of interest are silenced in Escherichia coli. The vectors are all IPTG-inducible and co-transformable in any combination and target genes are silenced conditionally and concurrently. Furthermore, in order to improve silencing efficacy, the vectors are designed to express uniquely shaped antisense RNAs, named paired termini antisense RNAs (PTasRNAs). The vectors are useful for comprehensive investigation of gene function and are applicable even if the target genes are essential for cell growth. Here, we describe methods to construct PTasRNA-expressing vectors and to evaluate silencing efficacy.

AB - We have developed four expression vectors to express antisense RNAs (asRNAs) by which genes of interest are silenced in Escherichia coli. The vectors are all IPTG-inducible and co-transformable in any combination and target genes are silenced conditionally and concurrently. Furthermore, in order to improve silencing efficacy, the vectors are designed to express uniquely shaped antisense RNAs, named paired termini antisense RNAs (PTasRNAs). The vectors are useful for comprehensive investigation of gene function and are applicable even if the target genes are essential for cell growth. Here, we describe methods to construct PTasRNA-expressing vectors and to evaluate silencing efficacy.

KW - Culture Techniques

KW - DNA, Bacterial

KW - DNA, Recombinant

KW - Escherichia coli

KW - Escherichia coli Proteins

KW - Gene Expression Regulation, Bacterial

KW - Gene Knockdown Techniques

KW - Gene Silencing

KW - Genetic Engineering

KW - Inverted Repeat Sequences

KW - Molecular Biology

KW - Molecular Sequence Data

KW - Polymerase Chain Reaction

KW - RNA, Antisense

KW - Transformation, Bacterial

KW - Journal Article

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

U2 - 10.1007/978-1-61779-424-7_23

DO - 10.1007/978-1-61779-424-7_23

M3 - Article

C2 - 22131001

SN - 1064-3745

VL - 815

SP - 307

EP - 319

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

Multiple-gene silencing using antisense RNAs in Escherichia coli

Abstract

Keywords

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