Part:BBa_K4274007

patB_target(gRNA)

Guide RNA (gRNA) is used as a molecule that helps with the cleavage of DNA since it guides the Cas nuclease to a targeted dsDNA sequence. As a gRNA, patB_target is used for knocking-out patB gene in the genome of Bacillus subtilis.

Usage and Biology

Single-guide RNA (sgRNA) is RNA designed artifially to be able to bind to a DNA sequence. It combines with the Cas9 protein to work to cleave the target RNA and though it is slightly shorter, it has the same function as the original RNAs in the CRISPR/Cas9 system. We designed the patB_target(gRNA) to specifically target the patB enzyme and to knock-out the gene in the the genome of Bacillus subtilis. This segment of RNA could also be used for gene-editing on the site of patB in Bacillus subtilis.

Source

Bacillus subtilis

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 183
  
• 1000
  COMPATIBLE WITH RFC[1000]

Reference

[1] Dang, Y., Jia, G., Choi, J. et al. Optimizing sgRNA structure to improve CRISPR-Cas9 knockout efficiency. Genome Biol 16 (4), 280 (2015). https://doi.org/10.1186/s13059-015-0846-3