Design and Preparation of a Multimeric Self-Cleaving Hammerhead Ribozyme
Open Access
- 1 February 1997
- journal article
- Published by Taylor & Francis in BioTechniques
- Vol. 22 (2) , 338-345
- https://doi.org/10.2144/97222rr03
Abstract
The activity of a ribozyme can be impaired by additional sequences at the 5′ and 3′ termini of the catalytic sequence. To approach this problem, a system was designed that minimizes sequences upstream and downstream from active regions of a hammerhead ribozyme and allows delivery of a large number of active molecules. A self-cleavable multimeric molecule was prepared by placing a ribozyme target sequence (derived from the core region of the hepatitis B virus [HBV]) upstream and downstream from the catalytic sequence. This construct was cloned in tandem into in vitro expression vectors. 32P-UTP-labeled transcripts of the multimeric construct, as well as non-self-cleaved monomeric ribozyme controls, and substrate were synthesized. The multimeric ribozyme molecule efficiently self-cleaved to release monomeric ribozymes lacking any extra upstream and downstream sequences. In addition, monomers were substantially more active against the HBV target RNA than the nonself- cleavable ribozymes. Up to 80% degradatio...Keywords
This publication has 0 references indexed in Scilit: