Part:BBa_K1797007

tet promoter-sgRNA-terminator

This part is modified from BBa_K1797005. We added promoter and terminator before and after the sgRNA for the convenience of users. Users can change the sequence of sgRNA(except for the sgRNA scaffold) for their own purpose.

Fig.1 The basic principles of CRIPSR/Cas9 system (source:http://blog.sciencenet.cn/blog-472747-723581.html)

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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 543
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 262
  Illegal BsaI site found at 440
  Illegal SapI site found at 84
  Illegal SapI.rc site found at 73