Target-Induced SOFA-HDV Ribozyme
Small cis-acting ribozymes have been converted into trans-acting ribozymes possessing the ability to cleave RNA substrates. The Hepatitis Delta Virus (HDV) ribozyme is one of the rare examples of these that is derived from an RNA species that is found in human cells. Consequently, it possesses the natural ability to function in the presence of human proteins in addition to an outstanding stability in human cells, two significant advantages in its use. The development of an additional specific on/off adaptor (SOFA) has led to the production of a new generation of HDV ribozymes with improved specificities that provide a tool with significant potential for future development in the fields of both functional genomics and gene therapy. SOFA-HDV ribozyme-based gene inactivation systems have been reported in both prokaryotic and eukaryotic cells. Here, a step-by-step approach for the efficient design of highly specific SOFA-HDV ribozymes with a minimum investment of time and effort is described.
Key wordsRibozyme Gene inactivation Functional genomics Gene therapy
We thank Jonathan Perreault and Gilles Boire for information about hY RNA. This work was supported by grants from Canadian Institute of Health Research (CIHR; grant numbers MOP-44002 and EOP-38322) to J.P.P. The RNA group is supported by grants both from CIHR and Université de Sherbrooke. M.V.L. was the recipient of a predoctoral fellowship from the Fonds de Recherche en Santé du Québec. J.P.P. holds the Canada Research Chairs in genomics and catalytic RNAs, and is a member of the Centre de Recherche Clinique Étienne Lebel.