Sequence‐dependent hydrolysis of RNA using modified oligonucleotide splints and RNase H

Abstract
We found that, in the presence of chimeric oligonucleotides containing complementary deoxyribo‐ and 2′‐O‐methylnucleosides, a nonaribonucleotide, [5′‐32 pACUUACCUG, was cleaved specifically upon treatment with RNase H. When 3′m(UG)d(AATG)m(GAC)5′ was used as a hybridization strand, *pACUUACCUG was cleaved between C6 and C7 to yield *pACUUAC. In the presence of 3′m(UGAA)d(TGGA)m(C)5′, the nonaribonucleotide was hydrolyzed, mainly between U8 and C9, to give *pACUUACCU. This method will have a variety of applications in the field of RNA engineering.