Difference between revisions of "Team:William and Mary/Description"
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<div><p>Once individual cell measurements have been taken for both CFP and YFP expression using confocal microscopy, the intrinsic noise measurements can be calculated (link to John’s part about this) and further analyzed.</p></div> | <div><p>Once individual cell measurements have been taken for both CFP and YFP expression using confocal microscopy, the intrinsic noise measurements can be calculated (link to John’s part about this) and further analyzed.</p></div> | ||
− | <div><p style="float: right;"><IMG SRC="https://static.igem.org/mediawiki/2015/9/97/WMdCasDiag.jpeg" width=600px></p><p>In addition to characterizing the noise of promoters in iGEM, we contributed additional tools for manipulating their expression. CRISPR/Cas9 has been used extensively in synthetic biology, both inside and out of iGEM. In particular, new functionalizations of the Cas9 protein that remove its catalytic nuclease domain while retaining its DNA-binding activity have allowed for novel methods in molecular biology. This catalytically inactive variant of Cas9, known as dCas9, can be used to repress gene expression by targeting the promoter region of a gene of interest. This repression is mediated by the CRISPR/Cas9 complex binding to the promoter region and can block RNAP binding or prevent transcriptional elongation. All of our gRNAs prevent RNAP binding and initiation of transcription.</p></div></p> | + | <div><p style="float: right;"><IMG SRC="https://static.igem.org/mediawiki/2015/9/97/WMdCasDiag.jpeg" width=600px></p><p> |
+ | Schematic of dCas9/CRISPR system. gRNAs targeting the promoter region of the gene of interest will guide dCas9 binding. Bound dCas9 sterically hinders RNAP and transcription factors from binding and promoting transcription.</p> | ||
+ | <p>In addition to characterizing the noise of promoters in iGEM, we contributed additional tools for manipulating their expression. CRISPR/Cas9 has been used extensively in synthetic biology, both inside and out of iGEM. In particular, new functionalizations of the Cas9 protein that remove its catalytic nuclease domain while retaining its DNA-binding activity have allowed for novel methods in molecular biology. This catalytically inactive variant of Cas9, known as dCas9, can be used to repress gene expression by targeting the promoter region of a gene of interest. This repression is mediated by the CRISPR/Cas9 complex binding to the promoter region and can block RNAP binding or prevent transcriptional elongation. All of our gRNAs prevent RNAP binding and initiation of transcription.</p></div></p> | ||
− | |||
− | |||
− | + | <div> | |
− | + | <p>References</p> | |
− | </p> | + | |
+ | <p>1: Elowitz, Michael B., et al. "Stochastic gene expression in a single cell." Science 297.5584 (2002): 1183-1186.</p> | ||
+ | |||
+ | <p>2: Raser, Jonathan M., and Erin K. O'Shea. "Noise in gene expression: origins, consequences, and control." Science 309.5743 (2005): 2010-2013. | ||
+ | </p> | ||
+ | </div> | ||
</div> | </div> | ||
</div> | </div> |
Revision as of 11:50, 18 September 2015
Integrator Suites
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Antibiotic Operons
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dCas9s
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gRNAs
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XFPs Under Various Promoters
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G^2
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