For gene expression, the coding sequences could be codon optimized and the level of gene expression is manageable by using different promoters and vector systems.

For gene silencing, we help the researchers to validate the potent shRNA.  The designed shRNAs are cloned into Biosettia’s lentiviral shRNA vectors and the corresponding target cDNA sequences are transcriptionally fused to the luciferase reporter to generate the target vectors; then the shRNA and target plasmids are co-transfected into the 293T cells.  The luciferase activity will be greatly reduced if the target : luciferase fusion gene expression is silenced by the potent shRNA(s) via the RNAi machinery in the 293T cells.

Gene Expression and Silencing

Diagram of a scientific process with labeled circular pathways, a sequence of green and yellow molecules, a DNA microarray scanner, a bar chart comparing reporter activity levels, and illustrations of enzyme and light reactions.

  • design 10 shRNA and clone into shRNA vector.

  • clone the shRNA target sequence fused with luciferase gene.

  • 293T co-transfection, screen the potent shRNA by luciferase reporter assay.

  • pick the most potent shRNA(s) for lentiviral production.

  • 5 x 1ml/vial per clone, titered 3E+7 to 1E+8 IU/ml.