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. 1995 Sep 11;23(17):3524–3530. doi: 10.1093/nar/23.17.3524

Double stranded scission of DNA directed through sequence-specific R-loop formation.

R Landgraf 1, C H Chen 1, D S Sigman 1
PMCID: PMC307233  PMID: 7567465

Abstract

R-loop formation with short (100 nt) RNAs provides a highly flexible and stringent method to achieve sequence-specific separation of target DNA at any given sequence. After stabilization of R-loops with glyoxal and removal of the RNA through RNase treatment the remaining single-stranded DNA bubble provides a highly favorable substrate for attenuated micrococcal nuclease. We investigated this method for sequence-specific scission of double-stranded DNA and achieved quantitative scission of 3-5 kb plasmids. The applicability to larger size DNA is demonstrated through specific excision of the intervening segment between two R-loops from a P1 plasmid of approximately 120 kb.

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Selected References

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