Difference between revisions of "Part:BBa K3044017"

Revision as of 16:44, 7 October 2019

sgRNA targeting GFP

This single guide RNA (sgRNA) sequence targets gene expression of mRFP (BB). 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 325-245 on the template strand in the mRFP sequence and have an efficiency of 67.06.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
 This single guide RNA (sgRNA) sequence targets gene expression of mRFP (BB).
-When utilizing the CRISPR/Cas system as a gene edition tool two components are essential; the Cas protein and the sgRNA. The Cas protein has endonuclease activity and induces double stranded DNA cleavage. The Cas protein forma a complex with the sgRNA, which serves as a guide for the Cas protein to ensure cleavage of the right target.
+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 Cas 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 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 Cas handle and interacts with the Cas protein.
+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 325-245 on the template strand in the mRFP sequence and have an efficiency of 67.06.