Protein Trans-splicing As a Means for Viral Vector-mediated in Vivo Gene Therapy

Overview

Journal Hum Gene Ther

Specialties Genetics
Pharmacology

Date 2008 Sep 16

PMID 18788906

Citations 59

Authors

Juan Li

Wunchang Sun

Bing Wang

Xiao Xiao

Xiang-Qin Liu

Affiliations

Soon will be listed here.

Abstract

Inteins catalyze protein splicing in a fashion similar to how self-splicing introns catalyze RNA splicing. Splitinteins catalyze precise ligation of two separate polypeptides through trans-splicing in a highly specific manner. Here we report a method of using protein trans-splicing to circumvent the packaging size limit of gene therapy vectors. To demonstrate this method, we chose a large dystrophin gene and an adeno-associated viral (AAV) vector, which has a small packaging size. A highly functional 6.3-kb Becker-form dystrophin cDNA was broken into two pieces and modified by adding appropriate split-intein coding sequences, resulting in splitgenes sufficiently small for packaging in AAV vectors. The two split-genes, after codelivery into target cells, produced two polypeptides that spontaneously trans-spliced to form the expected Becker-form dystrophin protein in cell culture in vitro. Delivering the split-genes by AAV1 vectors into the muscle of a mouse model of Duchenne muscular dystrophy rendered therapeutic gene expression and benefits.

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