Difference between revisions of "Part:BBa K4335013"
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− | The Cas9 protein is part of the bacteria's CRISPR/Cas9 immune defense mechanism to identify and destroy foreign DNA. By incorporating the foreign DNA into the bacteria's own DNA, it has a memory of any prior foreign DNA that the bacteria has encountered.The Cas9 endonuclease can be used either with the tracrRNA and crRNA (CRISPR RNA- Clustered Regularly Interspaced Short Palindromic Repeats) or with a guide RNA to cleave foreign DNA at a species locations. This makes exceptionally useful for DNA cloning in synthetic biology. | + | The Cas9 protein is part of the bacteria's CRISPR/Cas9 immune defense mechanism to identify and destroy foreign DNA. By incorporating the foreign DNA into the bacteria's own DNA, it has a memory of any prior foreign DNA that the bacteria has encountered.<br> |
+ | <br> | ||
+ | The Cas9 endonuclease can be used either with the tracrRNA and crRNA (CRISPR RNA- Clustered Regularly Interspaced Short Palindromic Repeats) or with a guide RNA to cleave foreign DNA at a species locations. This makes exceptionally useful for DNA cloning in synthetic biology. | ||
+ | <h2>Usage</h2> | ||
+ | We found a SpCas9 from iGEM16_Cambridge-JIC in the previous part <a href="https://parts.igem.org/Part:BBa_K2148013">[BBa_K2148013]</a> | ||
+ | .<br> | ||
+ | <br> | ||
+ | This part is based on the original Cas9 part in the registry, BBa_K1218011. To enhance the expression of Cas9 in the chloroplast chassis, Cambridge-JIC have codon-optimised it using the codon-optimisation software developed by Saul Purton.<br> | ||
+ | <br> | ||
+ | The amino acid and nucleotide sequences of BBa_K2148013 and BBa_K2148013 were compared. | ||
+ | <figure> | ||
+ | <img src="https://static.igem.wiki/teams/4335/wiki/cas9-1.jpg" width="100%" style="float:center"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:1rem">The comparison of nucleotide sequences of BBa_K2148013 and BBa_K4335013 showed that the matching rate was 67.96%. | ||
+ | </p> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <figure> | ||
+ | <img src="https://static.igem.wiki/teams/4335/wiki/cas9-2.png" width="100%" style="float:center"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:1rem">The amino acid sequences of BBa_K2148013 and BBa_K4335013 were compared and the matching degree was 100%. | ||
+ | </p> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | |||
+ | </html> | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== |
Revision as of 15:47, 10 October 2022
SpCas9
The Cas9 protein is part of the bacteria's CRISPR/Cas9 immune defense mechanism to identify and destroy foreign DNA. By incorporating the foreign DNA into the bacteria's own DNA, it has a memory of any prior foreign DNA that the bacteria has encountered.
The Cas9 endonuclease can be used either with the tracrRNA and crRNA (CRISPR RNA- Clustered Regularly Interspaced Short Palindromic Repeats) or with a guide RNA to cleave foreign DNA at a species locations. This makes exceptionally useful for DNA cloning in synthetic biology.
Usage
We found a SpCas9 from iGEM16_Cambridge-JIC in the previous part [BBa_K2148013] .This part is based on the original Cas9 part in the registry, BBa_K1218011. To enhance the expression of Cas9 in the chloroplast chassis, Cambridge-JIC have codon-optimised it using the codon-optimisation software developed by Saul Purton.
The amino acid and nucleotide sequences of BBa_K2148013 and BBa_K2148013 were compared. Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 241
Illegal PstI site found at 787
Illegal PstI site found at 2209
Illegal PstI site found at 2413
Illegal PstI site found at 2443
Illegal PstI site found at 3655 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 241
Illegal PstI site found at 787
Illegal PstI site found at 2209
Illegal PstI site found at 2413
Illegal PstI site found at 2443
Illegal PstI site found at 3655 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 248
Illegal BglII site found at 1043
Illegal BglII site found at 1733
Illegal BglII site found at 3812 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 241
Illegal PstI site found at 787
Illegal PstI site found at 2209
Illegal PstI site found at 2413
Illegal PstI site found at 2443
Illegal PstI site found at 3655 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 241
Illegal PstI site found at 787
Illegal PstI site found at 2209
Illegal PstI site found at 2413
Illegal PstI site found at 2443
Illegal PstI site found at 3655
Illegal NgoMIV site found at 1075 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 3468