Targeted cleavage of mRNA in vitro by RNase P from Escherichia coli

Y. Li, C. Guerrier-Takada, Sidney Altman

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

External guide sequences (EGSs) complementary to mRNAs that encode β- galactosidase from Escherichia coli and nuclease A from Staphylococcus aureus can target these RNAs for cleavage in vitro by RNase P from E. coli. Specific cleavage occurs at locations predicted by the nucleotide sequences of the EGSs. EGSs with regions complementary to the mRNAs that are as short as 13 nucleotides function efficiently and turn over slowly during incubation with the target substrate and the enzyme. EGSs composed of deoxyribonucleotides as well as those composed of ribonucleotides are effective, but cleavage of the targeted substrate with DNA as an EGS is about 10-fold less efficient than that with RNA as an EGS. An RNA EGS inhibited the formation of β- galactosidase activity in a crude extract (S30) of E. coli that was capable of catalyzing coupled transcription-translation reactions.

Original languageEnglish (US)
Pages (from-to)3185-3189
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number8
StatePublished - 1992
Externally publishedYes

Fingerprint

Ribonuclease P
Galactosidases
Escherichia coli
Messenger RNA
Guide RNA
RNA Cleavage
Deoxyribonucleotides
Ribonucleotides
Complex Mixtures
Staphylococcus aureus
Nucleotides
RNA
DNA
Enzymes
In Vitro Techniques

Keywords

  • external guide sequence
  • model substrates

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Targeted cleavage of mRNA in vitro by RNase P from Escherichia coli. / Li, Y.; Guerrier-Takada, C.; Altman, Sidney.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 8, 1992, p. 3185-3189.

Research output: Contribution to journalArticle

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