Trans-splicing ribozymes for targeted gene delivery Export

@article{kohler99, abstract = {Ribozymes are potential tools for genetic manipulation, and various naturally occurring catalytic RNAs have been dissected and used as the basis for the design of new endoribonuclease activities. While such cleaving ribozymes may work well in vitro, they have not proved to be routinely effective in depleting living cells of the chosen target RNA. Recently, trans-splicing ribozymes have been employed to repair mutant mRNAs in vivo. We have designed modified trans-splicing ribozymes with improved biological activity. These allow accurate splicing of a new 3' exon sequence into a chosen site within a target RNA, and in frame fusion of the exon can result in expression of a new gene product. These trans-splicing ribozymes contain catalytic sequences derived from a self-splicing group I intron, which have been adapted to a chosen target mRNA by fusion of a region of extended complementarity to the target RNA and precise alteration of the guide sequences required for substrate recognition. Both modifications are required for improved biological activity of the ribozymes. Whereas cleaving ribozymes must efficiently deplete a chosen mRNA species to be effective in vivo, even inefficient trans-splicing can allow the useful expression of a new gene activity, dependent on the presence of a chosen RNA. We have targeted trans-splicing ribozymes against mRNAs of chloramphenicol acetyltransferase, human immunodeficiency virus, and cucumber mosaic virus, and demonstrated trans-splicing and delivery of a marker gene in Escherichia coli cells. The improved trans-splicing ribozymes may be tailored for virtually any target RNA, and provide a new tool for triggering gene expression in specific cell types.}, author = {K\"{o}hler, Uwe and Ayre, Brian G. and Goodman, Howard M. and Haseloff, Jim}, citeulike-article-id = {2857581}, citeulike-linkout-0 = {http://dx.doi.org/10.1006/jmbi.1998.2447}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/9925776}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=9925776}, citeulike-linkout-3 = {http://www.sciencedirect.com/science/article/B6WK7-45R884M-RB/1/acc682a5186544113cd820a01c113259}, day = {5}, doi = {10.1006/jmbi.1998.2447}, journal = {Journal of Molecular Biology}, keywords = {design, genetherapy, ribozyme, tetrahymena, trans-splicing}, month = {February}, number = {5}, pages = {1935--1950}, posted-at = {2008-06-02 18:51:13}, priority = {0}, title = {Trans-splicing ribozymes for targeted gene delivery}, url = {http://dx.doi.org/10.1006/jmbi.1998.2447}, volume = {285}, year = {1999} }

TY - JOUR ID - kohler99 L3 - citeulike-article-id:2857581 N2 - Ribozymes are potential tools for genetic manipulation, and various naturally occurring catalytic RNAs have been dissected and used as the basis for the design of new endoribonuclease activities. While such cleaving ribozymes may work well in vitro, they have not proved to be routinely effective in depleting living cells of the chosen target RNA. Recently, trans-splicing ribozymes have been employed to repair mutant mRNAs in vivo. We have designed modified trans-splicing ribozymes with improved biological activity. These allow accurate splicing of a new 3' exon sequence into a chosen site within a target RNA, and in frame fusion of the exon can result in expression of a new gene product. These trans-splicing ribozymes contain catalytic sequences derived from a self-splicing group I intron, which have been adapted to a chosen target mRNA by fusion of a region of extended complementarity to the target RNA and precise alteration of the guide sequences required for substrate recognition. Both modifications are required for improved biological activity of the ribozymes. Whereas cleaving ribozymes must efficiently deplete a chosen mRNA species to be effective in vivo, even inefficient trans-splicing can allow the useful expression of a new gene activity, dependent on the presence of a chosen RNA. We have targeted trans-splicing ribozymes against mRNAs of chloramphenicol acetyltransferase, human immunodeficiency virus, and cucumber mosaic virus, and demonstrated trans-splicing and delivery of a marker gene in Escherichia coli cells. The improved trans-splicing ribozymes may be tailored for virtually any target RNA, and provide a new tool for triggering gene expression in specific cell types. IS - 5 JF - Journal of Molecular Biology EP - 1950 TI - Trans-splicing ribozymes for targeted gene delivery VL - 285 SP - 1935 KW - design KW - genetherapy KW - ribozyme KW - tetrahymena KW - trans-splicing AU - Köhler, Uwe AU - Ayre, Brian AU - Goodman, Howard AU - Haseloff, Jim PY - 1999/02/05/ UR - http://dx.doi.org/10.1006/jmbi.1998.2447 ER -