Site-specific cleavage of DNA-RNA hybrids by zinc finger/FokI cleavage domain fusions

Yang Gyun Kim, Yigong Shi, Jeremy M. Berg, Srinivasan Chandrasegaran

Research output: Contribution to journalArticle

Abstract

Zinc-finger proteins of the Cys2His2 type bind DNA-RNA hybrids with affinities comparable to those for DNA duplexes. Such zinc-finger proteins were converted into site-specific cleaving enzymes by fusing them to the FokI cleavage domain. The fusion proteins are active and under optimal conditions cleave DNA duplexes in a sequence-specific manner. These fusions also exhibit site-specific cleavage of the DNA strand within DNA-RNA hybrids albeit at a lower efficiency (≃50-fold) compared to the cleavage of the DNA duplexes. These engineered endonucleases represent the first of their kind in terms of their DNA-RNA cleavage properties, and they may have important biological applications.

Original languageEnglish (US)
Pages (from-to)43-49
Number of pages7
JournalGene
Volume203
Issue number1
DOIs
StatePublished - Dec 5 1997

Keywords

  • Chimeric restriction endonuclease
  • Flavorbacterium okeanokoites
  • Protein engineering
  • Recognition and cleavage domains

ASJC Scopus subject areas

  • Genetics

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