Difference between revisions of "Part:BBa K1431411:Design"
(→Design Notes) |
(→Design Notes) |
||
Line 9: | Line 9: | ||
But there are two problem: | But there are two problem: | ||
+ | |||
First,the DSB of binding sequence is hard to observe in the cell; | First,the DSB of binding sequence is hard to observe in the cell; | ||
+ | |||
Second,the whole HIV genome have a potential dangers in the lab. | Second,the whole HIV genome have a potential dangers in the lab. | ||
Revision as of 17:16, 16 October 2014
target sequence for HIV
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]
Design Notes
After we use design the gRNA of HIV-1(BBa_K1431401), we need the corresponding binding sequence DNA to test the efficiency of gRNA to lead a double strands break .
But there are two problem:
First,the DSB of binding sequence is hard to observe in the cell;
Second,the whole HIV genome have a potential dangers in the lab.
So we use the alternative solution.We insert the gRNA binding sequence between the promoter and reporter, transform the expression of DSB of HIV to the change of reporter. The whole target-sequence have a gRNA binding sequence in the middle and have more original sequence in two sides to return the natural state of HIV genome.
Source
HIV1_REF_2010_genome_DNA