Part:BBa_K3044012
sgRNA targeting GFP
This single guide RNA (sgRNA) sequence targets gene expression of GFP (BBa_E0040). When utilizing the CRISPR/Cas9 system as a gene edition tool two components are essential; the Cas9 protein and the sgRNA. The Cas9 protein has endonuclease activity and induces double stranded DNA cleavage. The Cas9 protein forms a complex with the sgRNA, which serves as a guide for the Cas9 protein to ensure cleavage of the right target. The Cas9 protein recognizes a PAM sequence and thereby knows there to cleave. The sgRNA is therefore, designed according to the location of the PAM sequences in the target gene. The sgRNA consists of 100 nucleotides of which 20 nucleotides basepair with the target DNA sequence and the rest is called a Cas9 handle and interacts with the Cas9 protein. This sgRNA sequence targets residue 250-269 on the template strand in the GFP sequence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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