Part:BBa_K1797006

dCas9

CRISPR Cas9 is a very popular gene editing system. This part includes an engineered dCas9 which is still able to bind to the DNA under the guidance of gRNA. However, it loses its catalytic activity. Actually, dCas9 has a very promising future. For example, We can use dCas9 to block the recruitment of transcription machine.

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.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1099
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 3378
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]