Difference between revisions of "Part:BBa K2927010"

Latest revision as of 05:27, 16 October 2019

DNA template of crRNA 1 for LbCas12a

This part encode the crRNA-1 that mediates Cas12a protein to recognize target sequences. Once crRNA binds with Cas12a protein, the complex capable to target and cleave targeted double-stranded DNA sequence (cis cleavage activity). The cleavage of double-stranded DNA activates the ability of Cas12a protein to cleave non-specific ssDNA (trans cleavage activity). In our project, crRNA-1 is designed to detect specific sequence of vp72 gene in African swine fever virus (ASFV).

Of note, crRNA-1 only contains 20 nucleotide sequences from vp72 gene of ASFV, which shows no potential of protein coding. Our design had been reported to iGEM and got approved.

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]

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 <partinfo>BBa_K2927010 short</partinfo>
-CrRNA is crucial for the CRISPR Cas system. It is the short sequence that combines with Cas12a protein and activates it. When crRNA binds on Cas12a protein, it makes Cas12a protein cleavage not only target sequence but also non-specific ssDNA. In our project, we use crRNA for detecting the African swine fever virus (ASFV). When crRNA 1 binds on Cas12a protein, the protein can not only detect the specific sequence of ASFV but also cleavage DNA of ASFV and non-specific ssDNA. By the way, though crRNA 1 is derived from ASFV, there is no possibility for virulence. The sequence of crRNA 1 is derived from p72 which is the capsid of ASFV. Therefore, the sequence of crRNA 1 is too short to become a functional protein. There is no concentration for spread out ASFV of our project.
+This part encode the crRNA-1 that mediates Cas12a protein to recognize target sequences. Once crRNA binds with Cas12a protein, the complex capable to target and cleave targeted double-stranded DNA sequence (cis cleavage activity). The cleavage of double-stranded DNA activates the ability of Cas12a protein to cleave non-specific ssDNA (trans cleavage activity). In our project, crRNA-1 is designed to detect specific sequence of vp72 gene in African swine fever virus (ASFV).
+Of note, crRNA-1 only contains 20 nucleotide sequences from vp72 gene of ASFV, which shows no potential of protein coding.  Our design had been reported to iGEM and got approved.
+&#8195;
 <!-- Add more about the biology of this part here