Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs

P. A. Krieg; D. A. Melton

doi:10.1093/nar/12.18.7057

Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs

P. A. Krieg, D. A. Melton

Cellular and Molecular Medicine

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1286 Scopus citations

Abstract

We describe a method for the synthesis of microgram quantities of eucaryotic messenger RNAs. Injection into the cytoplasm of frog oocytes and addition to wheat germ extracts show that these synthetic RNAs function efficiently as messenger RNAs. We confirm that a 5′ cap on the mRNA is essential for translation in injected oocytes and show that most of the 3′ flanking region, including the poly A tail, can be deleted without the abolition of protein synthesis. The method of mRNA synthesis involves in vitro transcription of cDNAs which have been cloned into SP6 vectors (described in the accompanying paper). This method enables one to produce large amounts of mRNA and consequently protein from any cDNA clone.

Original language	English (US)
Pages (from-to)	7057-7070
Number of pages	14
Journal	Nucleic acids research
Volume	12
Issue number	18
DOIs	https://doi.org/10.1093/nar/12.18.7057
State	Published - Sep 25 1984

ASJC Scopus subject areas

Genetics

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title = "Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs",

abstract = "We describe a method for the synthesis of microgram quantities of eucaryotic messenger RNAs. Injection into the cytoplasm of frog oocytes and addition to wheat germ extracts show that these synthetic RNAs function efficiently as messenger RNAs. We confirm that a 5′ cap on the mRNA is essential for translation in injected oocytes and show that most of the 3′ flanking region, including the poly A tail, can be deleted without the abolition of protein synthesis. The method of mRNA synthesis involves in vitro transcription of cDNAs which have been cloned into SP6 vectors (described in the accompanying paper). This method enables one to produce large amounts of mRNA and consequently protein from any cDNA clone.",

author = "Krieg, {P. A.} and Melton, {D. A.}",

note = "Funding Information: ACKNOWLEDGEMENTS We thank J. Williams for supplying Xenopus globin DNAs and D. Falb for assistance in some plasmid constructions. We are grateful to H. Perry, M. Rebagliati, and D. Weeks for their comments on the manuscript and to K. Breakey for preparing the figures. P.K. acknowledges support from the Fogarty International Fellowship Program. This work was supported by grants from the N1H and The Chicago Community Trust/Searle Scholars Program to D.M.",

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AU - Krieg, P. A.

AU - Melton, D. A.

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PY - 1984/9/25

Y1 - 1984/9/25

N2 - We describe a method for the synthesis of microgram quantities of eucaryotic messenger RNAs. Injection into the cytoplasm of frog oocytes and addition to wheat germ extracts show that these synthetic RNAs function efficiently as messenger RNAs. We confirm that a 5′ cap on the mRNA is essential for translation in injected oocytes and show that most of the 3′ flanking region, including the poly A tail, can be deleted without the abolition of protein synthesis. The method of mRNA synthesis involves in vitro transcription of cDNAs which have been cloned into SP6 vectors (described in the accompanying paper). This method enables one to produce large amounts of mRNA and consequently protein from any cDNA clone.

AB - We describe a method for the synthesis of microgram quantities of eucaryotic messenger RNAs. Injection into the cytoplasm of frog oocytes and addition to wheat germ extracts show that these synthetic RNAs function efficiently as messenger RNAs. We confirm that a 5′ cap on the mRNA is essential for translation in injected oocytes and show that most of the 3′ flanking region, including the poly A tail, can be deleted without the abolition of protein synthesis. The method of mRNA synthesis involves in vitro transcription of cDNAs which have been cloned into SP6 vectors (described in the accompanying paper). This method enables one to produce large amounts of mRNA and consequently protein from any cDNA clone.

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ER -

Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs

Abstract

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