Hi Mike & all, Great thread. I just wanted to drop in and make a contribution. Here is an interesting paper...if they can get it to work efficiently in vivo then there is a new player in town(imo)I don't think they are public though :(. Best of Luck, Where'd He Go?
ncbi.nlm.nih.gov
Nat Biotechnol 1999 Mar;17(3):246-52
Spliceosome-mediated RNA trans-splicing as a tool for gene therapy.
Puttaraju M, Jamison SF, Mansfield SG, Garcia-Blanco MA, Mitchell LG
Intronn LLC, Durham, NC 27701, USA.
[Medline record in process]
We have developed RNA molecules capable of effecting spliceosome-mediated RNA trans-splicing reactions with a target messenger RNA precursor (pre-mRNA). Targeted trans-splicing was demonstrated in a HeLa nuclear extract, cultured human cells, and H1299 human lung cancer tumors in athymic mice. Trans-splicing between a cancer-associated pre-mRNA encoding the beta-subunit of human chorionic gonadotropin gene 6 and pre-trans-splicing molecule (PTM) RNA was accurate both in vitro and in vivo. Comparison of targeted versus nontargeted trans-splicing revealed a moderate level of specificity, which was improved by the addition of an internal inverted repeat encompassing the PTM splice site. Competition between cis- and trans-splicing demonstrated that cis-splicing can be inhibited by trans-splicing. RNA repair in a splicing model of a nonfunctional lacZ transcript was effected in cells by a PTM, which restored significant beta-galactosidase activity. These observations suggest that spliceosome-mediated RNA trans-splicing may represent a general approach for reprogramming the sequence of targeted transcripts, providing a novel approach to gene therapy.
PMID: 10096291, UI: 99193955 |
