RNA
Part:BBa_K1431403:Design
Designed by: Fan Jiang, Peng Peng Group: iGEM14_SUSTC-Shenzhen (2014-10-14)
gRNA2 for HBV
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Designing the Sequence
We used a method derived from the method described in the paper by Feng Zhang[http://www.nature.com/nbt/journal/v31/n9/abs/nbt.2647.html ZhangFgRNA].
The whole process can be divided into the following steps:
Conserved Sequence Analysis
We first extracted all conserved regions from the NIH HIV-1 Reference Genome. In this step, we found around 10 alternatives for the next process. Here all screening processes are done in a per-strain basis because of the high mutability of the HIV-1 virus.
Strip out sequences without PAM
Select gRNA sequences with the best theoretical quality
Source
AB010289-AB078031 in HBV_aligned Database