Part:BBa_K1797005
sgRNA
This part includes sgRNA, which can be used to do site-specific gene editing when CRISPR/Cas9 system is present. The sgRNA changeable region can be changed due to the purpose of uses. Users can use BspQI to digest the part and ligate with a DNA sequence that has BspQI compatible ends. The DNA sequence is designed according to the gene sequence that needs gene editing.
To make this part more clear, we find it necessary to introduce some background knowledge about CRIPSR/Cas9 gene-editing system. CRISPR/Cas9 used to be a bacterial adaptive immune system. When phages or plasmids get into the cell, this system will cut the DNA and store a part of the sequence in the host genome. When next time some exogenous DNA containing the stored DNA invades, the gRNA(guide RNA) and another RNA molecule essential for the association between gRNA and Cas9 protein will be transcribed, guiding Cas9 protein to the gRNA targeted locus. Cas9 protein is a DNA endonuclease, causing DSB(double-stranded break) in the targeted locus. DSB initiates homologous recombination(HR) or non homologous end joining(NHEJ) depending on whether there is a homologous fragment or not. Researchers around the world are using this technology to do site-specific gene editing. In addition, the rapidly developing technologies make the combination of RNA molecules into sgRNA(single guide RNA) involved in this process come true, which significantly improves the convenience of using this gene-editing system. Here we introduce the sgRNA into the iGEM Part Registry to help those iGEM teams in need.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 98
Illegal BamHI site found at 650 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 666
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 369
Illegal BsaI site found at 547
Illegal SapI site found at 191
Illegal SapI.rc site found at 180
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