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

• Published in: FEBS letters Vol. 215; no. 2; pp. 327 - 330
• Main Authors: Inoue, Hideo, Hayase, Yoji, Iwai, Shigenori, Ohtsuka, Eiko
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 11-05-1987; Elsevier
• Subjects: 2′- O-Methyloligonucleotide; Analytical, structural and metabolic biochemistry; Autoradiography; Base Sequence; Biological and medical sciences; Chimeric oligonucleotide; Endoribonucleases; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Heteroduplex; Hydrolases; Hydrolysis; Nucleic Acid Hybridization; Oligonucleotides; Ribonuclease H; RNA; RNA hydrolysis; RNase H; Site-specific cleavage; Heteroduplex; 2′- O-Methyloligonucleotide; Site-specific cleavage; RNase H; Chimeric oligonucleotide; RNA hydrolysis; Hydrolysis; RNA; Enzyme; Reaction mechanism; Ribonuclease H; DNA RNA hybrid; Oligodeoxyribonucleotide
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(87)80171-0

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.