Part:BBa_K3885172

Guide RNA (gRNA) target for tetR

Guide RNA (gRNA) is a piece of RNAs that function as guides for RNA- or DNA-targeting enzymes, which they form complexes with. Very often these enzymes will delete, insert or otherwise alter the targeted DNA.

Usage and Biology

Figure 1. Schematic of the design procedure of the gRNA construction and screening. There are five 20nt sequences to change into five different gRNAs.

In the CRISPR/Cas9 system, gRNA can direct Cas9 protein to cut dsDNA at the sequence target site. Simultaneously, a hybridization region of igRNA is adding additional sequences in the 5' of sgRNA. Therefore, designing an active gRNA with high cleavage ability is important.
Based on the target gene(tetR), we chose CGG and AGG as PAM, and designed 5 gRNAs in the plasmid pTargetF (Figure 1).

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]