Difference between revisions of "Part:BBa K1633007:Design"
(→Design Notes) |
|||
| Line 9: | Line 9: | ||
We designed specific GFP siRNAs based on a free software accessible online. This tool can find the best siRNA sequences on target gene GFP 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. | We designed specific GFP siRNAs based on a free software accessible online. This tool can find the best siRNA sequences on target gene GFP 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. | ||
| − | + | This part is artificial designed to target and downregulate GFP protein. This part is a short hairpin RNA (shRNA) sequence. When this shRNA sequence is cut by restriction enzyme and then integrated into pcDNA 6.2 vector, this shRNA can play a RNAi function in mammalian cell lines such as HEK293 cell. When the shRNA vector of GFP is transfected into mammalian cells, the shRNA hairpin structure is cleaved by Dicer into siRNA of GFP and loaded into the RISC. The siRNA-RISC complex targets at GFP mRNA under the guide of siRNA sequence and cleave the GFP mRNA. | |
| − | + | ||
===Source=== | ===Source=== | ||
Revision as of 12:09, 18 September 2015
GFP siRNA
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We designed specific GFP siRNAs based on a free software accessible online. This tool can find the best siRNA sequences on target gene GFP 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.
This part is artificial designed to target and downregulate GFP protein. This part is a short hairpin RNA (shRNA) sequence. When this shRNA sequence is cut by restriction enzyme and then integrated into pcDNA 6.2 vector, this shRNA can play a RNAi function in mammalian cell lines such as HEK293 cell. When the shRNA vector of GFP is transfected into mammalian cells, the shRNA hairpin structure is cleaved by Dicer into siRNA of GFP and loaded into the RISC. The siRNA-RISC complex targets at GFP mRNA under the guide of siRNA sequence and cleave the GFP mRNA.
Source
We designed the GFP 122 siRNA and ordered the part from a DNA synthesis company.