In vitro coupled transcription splicing

Barbara J. Natalizio; Mariano A. Garcia-Blanco

doi:10.1016/j.ymeth.2005.07.013

In vitro coupled transcription splicing

Barbara J. Natalizio, Mariano A. Garcia-Blanco

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

13 Scopus citations

Abstract

Many convincing studies published in recent years strongly support coupling of transcription and pre-mRNA processing. Despite key advances in our understanding of these processes, there is a lack of a robust in vitro system in which to study the mechanism of this coupling for complex pre-mRNAs. Here, we describe an in vitro system capable of transcribing and splicing complex transcripts with three and four exons. We also demonstrate how the system can be used to study exon silencing in vitro. We believe that this system will be a useful tool to study the mechanisms that mediate the coupling of transcription and pre-mRNA processing.

Original language	English (US)
Pages (from-to)	314-322
Number of pages	9
Journal	Methods
Volume	37
Issue number	4
DOIs	https://doi.org/10.1016/j.ymeth.2005.07.013
State	Published - Dec 2005
Externally published	Yes

Keywords

Cotranscriptional splicing
Coupling
RNA polymerase II
T7 RNA polymerase
Transcription
mRNA processing

ASJC Scopus subject areas

Molecular Biology
General Biochemistry, Genetics and Molecular Biology

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10.1016/j.ymeth.2005.07.013

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title = "In vitro coupled transcription splicing",

abstract = "Many convincing studies published in recent years strongly support coupling of transcription and pre-mRNA processing. Despite key advances in our understanding of these processes, there is a lack of a robust in vitro system in which to study the mechanism of this coupling for complex pre-mRNAs. Here, we describe an in vitro system capable of transcribing and splicing complex transcripts with three and four exons. We also demonstrate how the system can be used to study exon silencing in vitro. We believe that this system will be a useful tool to study the mechanisms that mediate the coupling of transcription and pre-mRNA processing.",

keywords = "Cotranscriptional splicing, Coupling, RNA polymerase II, T7 RNA polymerase, Transcription, mRNA processing",

author = "Natalizio, {Barbara J.} and Garcia-Blanco, {Mariano A.}",

note = "Funding Information: We thank all members of the Garcia-Blanco laboratory, especially James Pearson, for all the invaluable advice and helpful suggestions and Klemens Hertel for generously sharing data before publication and for his comments on this article. We thank Bruce Sullenger and Bryan Cullen for their generous gifts of the β-globin plasmids. This work was funded by NIH Grant RO1 GM071037 to M.G.B.",

year = "2005",

month = dec,

doi = "10.1016/j.ymeth.2005.07.013",

language = "English (US)",

volume = "37",

pages = "314--322",

journal = "Methods",

issn = "1046-2023",

publisher = "Academic Press Inc.",

number = "4",

}

TY - JOUR

T1 - In vitro coupled transcription splicing

AU - Natalizio, Barbara J.

AU - Garcia-Blanco, Mariano A.

N1 - Funding Information: We thank all members of the Garcia-Blanco laboratory, especially James Pearson, for all the invaluable advice and helpful suggestions and Klemens Hertel for generously sharing data before publication and for his comments on this article. We thank Bruce Sullenger and Bryan Cullen for their generous gifts of the β-globin plasmids. This work was funded by NIH Grant RO1 GM071037 to M.G.B.

PY - 2005/12

Y1 - 2005/12

N2 - Many convincing studies published in recent years strongly support coupling of transcription and pre-mRNA processing. Despite key advances in our understanding of these processes, there is a lack of a robust in vitro system in which to study the mechanism of this coupling for complex pre-mRNAs. Here, we describe an in vitro system capable of transcribing and splicing complex transcripts with three and four exons. We also demonstrate how the system can be used to study exon silencing in vitro. We believe that this system will be a useful tool to study the mechanisms that mediate the coupling of transcription and pre-mRNA processing.

AB - Many convincing studies published in recent years strongly support coupling of transcription and pre-mRNA processing. Despite key advances in our understanding of these processes, there is a lack of a robust in vitro system in which to study the mechanism of this coupling for complex pre-mRNAs. Here, we describe an in vitro system capable of transcribing and splicing complex transcripts with three and four exons. We also demonstrate how the system can be used to study exon silencing in vitro. We believe that this system will be a useful tool to study the mechanisms that mediate the coupling of transcription and pre-mRNA processing.

KW - Cotranscriptional splicing

KW - Coupling

KW - RNA polymerase II

KW - T7 RNA polymerase

KW - Transcription

KW - mRNA processing

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DO - 10.1016/j.ymeth.2005.07.013

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SN - 1046-2023

VL - 37

SP - 314

EP - 322

JO - Methods

JF - Methods

IS - 4

ER -

In vitro coupled transcription splicing

Abstract

Keywords

ASJC Scopus subject areas

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