Part:BBa_K2208005:Design
A shRNA corresponding DNA sequence for BCL2 which could silence the gene.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
RNA interference (RNAi) serves as a powerful tool that employs siRNAs to silence the expression of specific target genes. We designed specific BCL2 siRNAs to act as therapeutic agents to degrade BCL2 mRNA and block BCL2 expression and therefore allow higher apoptosis probabilities. Four siRNA sequences targeting different sites of the BCL2 mRNA open reading frame (ORF) were designed based on a free software accessible online. We conducted pre-experiments to find the best siRNA sequences on the targeted gene BCL2 to insure the maximum gene-specificity and silencing efficacy. This tool also designs the pair of oligonucleotides needed to generate short hairpin RNAs (shRNAs) in the plasmid. At least two of the four pre-designed shRNA plasmids are guaranteed to knock down expression of the targeted gene BCL2. The access to two effective sequences allows appropriate control on non-specific effects. When the shRNA plasmids of BCL2 are transfected into HEK293 cells, Dicer cleaves the shRNA into siRNA of BCL2. However, because siRNAs in vivo are vulnerable to degradation by plasma and tissue nucleases, nano-vesicles to facilitate siRNA uptake into the neurons need to be established.
Design Notes
We built the plasmid based on the building rules such as making sure the plasmids are chlorampenicol resistant and they hold restriction enzyme cutting site and so on.
Source
Artificially designed and synthesized by GenScript Company.