Publications [#99801] of Bruce A Sullenger

Papers Published

1. CS Rogers, CG Vanoye, BA Sullenger, AL George Jr, Functional repair of a mutant chloride channel using a trans-splicing ribozyme., The Journal of clinical investigation, vol. 110 no. 12 (December, 2002), pp. 1783-9, ISSN 0021-9738 [doi]
   (last updated on 2013/05/16)

   Abstract:
   RNA repair has been proposed as a novel gene-based therapeutic strategy. Modified Tetrahymena group I intron ribozymes have been used to mediate trans-splicing of therapeutically relevant RNA transcripts, but the efficiency of the ribozyme-mediated RNA repair process has not been determined precisely and subsequent restoration of protein function has been demonstrated only by indirect means. We engineered a ribozyme that targets the mRNA of a mutant canine skeletal muscle chloride channel (cClC-1) (mutation T268M in ClC-1 causing myotonia congenita) and replaces the mutant-containing 3' portion by trans-splicing the corresponding 4-kb wild-type sequence. Repair efficiency assessed by quantitative RT-PCR was 1.2% +/- 0.1% in a population of treated cells. However, when chloride channel function was examined in single cells, a wide range of electrophysiological activity was observed, with 18% of cells exhibiting significant functional restoration and some cells exhibiting complete rescue of the biophysical phenotype. These results indicate that RNA repair can restore wild-type protein activity and reveal considerable cell-to-cell variability in ribozyme-mediated trans-splicing reaction efficiency.

   Keywords:
   Animals • Base Sequence • Cell Line • Chloride Channels • Dogs • Exons • Humans • Molecular Sequence Data • Muscle, Skeletal • Mutation* • Myotonia Congenita • Patch-Clamp Techniques • Protozoan Proteins • RNA, Catalytic • RNA, Messenger • Reverse Transcriptase Polymerase Chain Reaction • Tetrahymena • Trans-Splicing* • genetics • genetics* • metabolism • metabolism*