Part:BBa_K4687013:Design
gRNA-(PAM/TTTG(-))
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The guide sequences in the gRNAs are paired with each other and the specific DNA sequences of the target gene or genome to ensure that the CRISPR-MAD7 complexes can accurately recognize and bind to the target DNA and cleave the DNA through the nuclease activity of the Cas proteins, thus completing the gene editing. In order to better improve the gene editing efficiency of the CRISPR-MAD7 system in Corynebacterium glutamicum during the experiment, we identified different PAM sites by designing different gRNAs, and determined the PAM site with the most prominent efficiency.
Source
The gRNAs are mainly designed based on the gene sequence of the PAM site. The sequence design mainly relies on the neighboring motifs located in the sequence of the pre-interval region of the PAM sequence (Protospacer Adjacent Motif).