The Effect of Cassette Sequence on the Knockdown Efficiency of a Triple tRNA-shRNA Chimera Gene Therapy Vector
Authors:Karl Haushalter, fang-chu lin
Mentor:Karl Haushalter, Associate Professor of Chemistry and Biology , Harvey Mudd College
In our lab, we are trying to find a safe and effective gene therapy for HIV-AIDS. A proposed method is to knockdown the human cellular protein, CCR5, which is a co-receptor for HIV entry into the cell. Using RNA interference is one of the potential strategies for gene therapy, and previous study in our lab found out that short hairpin RNA offers the best structure with this technique. So, for the chimera we used in our system, we have shRNAs fused to their respective tRNAs. Also, previous studies in our lab show that only minor differences in knockdown efficiency by other shRNAs expressed in the same vector, demonstrating that the individual cassettes act largely independently, and the use of a triple chimera construct did not significantly affect CCR5 knockdown activity compared to a single chimera construct. Therefore, we tried to change the sequence of three tRNA-shRNAs in a triple chimera construct to investigate how this new sequence would affect the CCR5 knockdown activity.