Small packaging capacity (less than 5kb) has limited the application of AAV in delivery of large genes. ViGene Biosciences provides trans-splicing and Cre dependent approach to overcome packaging restraints. Multiple AAV vectors coinfection can get into one cell at very high efficiency (more than 90%). Large gene can be split to two parts and the expression of whole gene can be reconstituted in vivo through the recombination of ITR, and right protein expression can be achieved by mRNA splicing. In order to decrease the inhibitory effects of ITR in transcription, Cre-LoxP can be used to eliminate ITR sequence. When compared to single vector, trans-splicing with Cre is about 20% expression efficiency.
View larger image. Two vector trans-splicing system
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