Difference between revisions of "Part:BBa K1431411:Design"

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===Design Notes===
 
===Design Notes===
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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 .
+
After we use design the gRNA of HIV-1([https://parts.igem.org/Part:BBa_K1431401 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:
 
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.
  
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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.
+
So we use the alternative solution.We insert the gRNA binding sequence between the promoter and reporter, transform the expression of DSB of binding sequence 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.
 
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===
 
===Source===
  
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HIV1_REF_2010_genome_DNA
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Conserved Region of the HIV-1 Genome from the NIH HIV-1 Sequence Database
  
 
===References===
 
===References===

Latest revision as of 03:48, 18 October 2014

target sequence for HIV


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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 binding sequence 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

Conserved Region of the HIV-1 Genome from the NIH HIV-1 Sequence Database

References