Difference between revisions of "Team:Edinburgh OG/Basic Part"

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<h3> Table of the Basic Parts BioBricks</h3>
 
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<tr><th style="width:8px" class="header"></th><th style="width:8px" class="header"></th><th style="width:8px" class="header"></th><th style="width: 87px;" class="header">Name</th><th style="width: 80px" colspan="" class="header">Type</th><th style="width: auto;" colspan="" class="header">Description</th><th style="width: 150px" colspan="" class="header">Designer</th><th style="width: 50px" colspan="" class="header">Length</th>
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<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_green">W</td><td><a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2330000">BBa_K2330000</a></td><td>Coding</td><td>SaCas9 codon-optimised for E. coli</td><td width="100px">filippo abbondanza</td><td align="right">3183</td>
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            <td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2330000">BBa_K2330000</a></td>
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            <td>Coding</td>    
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            <td>SaCas9 codon-optimised for E. coli</td>
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<groupparts>iGEM17 Edinburgh_OG</groupparts>
 
<groupparts>iGEM17 Edinburgh_OG</groupparts>
  

Revision as of 02:36, 2 November 2017

PhagED: a molecular toolkit to re-sensitise ESKAPE pathogens

Basic 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.

Table of the Basic Parts BioBricks

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NameTypeDescriptionDesignerLength
 W<a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2330000">BBa_K2330000</a>CodingSaCas9 codon-optimised for E. colifilippo abbondanza3183

<groupparts>iGEM17 Edinburgh_OG</groupparts>

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.