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<h1 class="subtitle text-center">Parts</h1> | <h1 class="subtitle text-center">Parts</h1> | ||
<p>This year the iGEM Edinburgh_OG team focused on developing a modular toolkit using CRISPR systems and phages to re-sensitise antibiotic-resistant bacteria. As a BioBrick we submit the <em>E. coli </em>codon-optimised <em>Staphylococcus aureus Cas9</em>.</p> | <p>This year the iGEM Edinburgh_OG team focused on developing a modular toolkit using CRISPR systems and phages to re-sensitise antibiotic-resistant bacteria. As a BioBrick we submit the <em>E. coli </em>codon-optimised <em>Staphylococcus aureus Cas9</em>.</p> | ||
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<groupparts>iGEM17 Edinburgh_OG</groupparts> | <groupparts>iGEM17 Edinburgh_OG</groupparts> | ||
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<div class="div-fig"> | <div class="div-fig"> | ||
<img src="https://static.igem.org/mediawiki/2017/c/c6/T--Edinburgh_OG--cas.png"> | <img src="https://static.igem.org/mediawiki/2017/c/c6/T--Edinburgh_OG--cas.png"> | ||
<p>http://www.nature.com/news/genome-editing-revolution-my-whirlwind-year-with-crispr-1.19063</p> | <p>http://www.nature.com/news/genome-editing-revolution-my-whirlwind-year-with-crispr-1.19063</p> | ||
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<h2>How does this part work?</h2> | <h2>How does this part work?</h2> | ||
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<li>To programme SaCas9, you need to design guide RNA (tracrRNA [2], 21 bp spacer flanked by direct repeats [2] ).</li> | <li>To programme SaCas9, you need to design guide RNA (tracrRNA [2], 21 bp spacer flanked by direct repeats [2] ).</li> | ||
</ul> | </ul> | ||
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<h3>Advantages of SaCas9 compared with the conventional <em>Streptococcus pyogenes</em> Cas9:</h3> | <h3>Advantages of SaCas9 compared with the conventional <em>Streptococcus pyogenes</em> Cas9:</h3> | ||
<ul> | <ul> | ||
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<li>Higher efficiency of SaCas9 over SpCas9 [2]</li> | <li>Higher efficiency of SaCas9 over SpCas9 [2]</li> | ||
</ul> | </ul> | ||
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<div class="div-ref"> | <div class="div-ref"> | ||
<p>[1] Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche, B., Shalem, O., Wu, X., Makarova, K. S., Koonin, E. V. Sharp, P.A., Zhang, F. 2015. In vivo genome editing using Staphylococcus aureus Cas9. Nature. 520 (7546). | <p>[1] Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche, B., Shalem, O., Wu, X., Makarova, K. S., Koonin, E. V. Sharp, P.A., Zhang, F. 2015. In vivo genome editing using Staphylococcus aureus Cas9. Nature. 520 (7546). |
Revision as of 01:11, 30 October 2017
Parts
This year the iGEM Edinburgh_OG team focused on developing a modular toolkit using CRISPR systems and phages to re-sensitise antibiotic-resistant bacteria. As a BioBrick we submit the E. coli codon-optimised Staphylococcus aureus Cas9.
<img src="">
http://www.nature.com/news/genome-editing-revolution-my-whirlwind-year-with-crispr-1.19063
How does this part work?
- Our SaCas9 can be programmed to cleave specific target sequence followed by the PAM sequence (5’-NNGRRT-3’).
- To express SaCas9, it requires suitable machinery such as promoter, RBS,and terminator.
- To programme SaCas9, you need to design guide RNA (tracrRNA [2], 21 bp spacer flanked by direct repeats [2] ).
Advantages of SaCas9 compared with the conventional Streptococcus pyogenes Cas9:
- Smaller size (1053 amino acids against 1368) resulting in an easier expression/delivery
- Different PAM sequence recognised (5’-NNGRRT-3’ ) increasing the usability
- Higher efficiency of SaCas9 over SpCas9 [2]
[1] Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche, B., Shalem, O., Wu, X., Makarova, K. S., Koonin, E. V. Sharp, P.A., Zhang, F. 2015. In vivo genome editing using Staphylococcus aureus Cas9. Nature. 520 (7546). pp.186-191.
[2] Friedland AE, Baral R, Singhal P, et al. Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications. Genome Biology. 2015;16:257. doi:10.1186/s13059-015-0817-8.
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