University Patents, Inc.

Competitive Technologies, Inc.

Other

Arthur J Zaug

Thomas R Cech

Michael D Been

Cheryl Grosshans

Felicia L Murphy

RNA enzymes or ribozymes can act as endoribonucleases, catalyzing the cleavage of RNA molecules with a sequence specificity of cleavage greater than that of known ribonucleases and approaching that of the DNA restriction endonucleases, thus serving as RNA sequence specific endoribonucleases. An example is a shortened form of the self-splicing ribosomal RNA intervening sequence of Tetrahymena (L-19 IVS RNA). Site-specific mutagenesis of the enzyme active site of the L-19 IVS RNA alters the substrate sequence specificity in a predictable manner, allowing a set of sequence-specific endoribonucleases to be synthesized. Varying conditions allow the ribozyme to act as a polymerase (nucleotidyltransferase), a dephosphorylase (acid phosphatase or phosphotransferase) or a sequence-specific endoribonuclease.

RNA ribozyme polymerases, dephosphorylases, restriction endoribonucleases and methods

RNA ribozyme polymerases, dephosphorylases, restriction

RNA Ribozyme which cleaves substrate RNA without formation of a

New RNA endoribonuclease ribozymes are found with new conditions to prevent mismatch cleavage and able to cleave RNA after 6 different sets of ribonucleotide 4 base sequences.

RNA ribozyme restriction endoribonucleases and methods

RNA enzymes or ribozymes can act as endoribonucleases, catalyzing the cleavage of RNA molecules with a sequence specificity of cleavage greater than that of known ribonucleases and approaching that of the DNA restriction endonucleases, thus serving as RNA sequence specific endoribonucleases. An example is a shortened form of the self-splicing ribonsomal RNA intervening sequence of Tetrahymena (L-19 IVS RNA). Site-specific mutagenesis of the enzyme active site of the L-19 IVS RNA alters the substrate sequence specificity in a predictable manner, allowing a set of sequence-specific endoribonucleases to be synthesized. Varying conditions allow the ribozyme to act as a polymerase (nucleotidyltransferase), a dephosphorylase (acid phosphatase or phosphotransferase) or a sequence-specific endoribonuclease.

A catalytic RNA (ribozyme) derived from an intervening sequence (IVS) RNA of Tetrahymena thermophila will catalyze an RNA polymerization reaction in which pentacytidylic acid (C.sub.5) is extended by the successive addition of mononucleotides derived from a guanylyl-(3',5')-nucleotide (GpN). Cytidines or uridines are added to C.sub.5 to generate chain lengths of 10 to 11 nucleotides; longer products are also generated but at reduced efficiency. The reaction is analogous to that catalyzed by a replicase with C.sub.5 acting as the primer, GpNs as the nucleoside triphosphates, and a sequence in the ribozyme providing a template.

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Louisville, CO, United States of America

Arthur J Zaug