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Team:SJTU-BioX-Shanghai/Prospect
From 2013.igem.org
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=CRISPRi-on= | =CRISPRi-on= | ||
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At present, our Metabolic Gear Box, which combines CRISPRi with light sensors, only down-regulates three genes of a pathway. However, in many other cases, genes in a synthetic pathway are supposed to be up-regulated in order to acquire the most products. | At present, our Metabolic Gear Box, which combines CRISPRi with light sensors, only down-regulates three genes of a pathway. However, in many other cases, genes in a synthetic pathway are supposed to be up-regulated in order to acquire the most products. | ||
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A newly devised tool, CRISPR-on, has provided some inception. CRISPR-on devisers simply fuse a transcriptional activation domain with dCas9 to create a sequence-specific transcription activating tool. CRISPR-on has been proved to be effective in human and mouse. We expect to conduct similar work in our bio-factory, ''Escherichia coli''. | A newly devised tool, CRISPR-on, has provided some inception. CRISPR-on devisers simply fuse a transcriptional activation domain with dCas9 to create a sequence-specific transcription activating tool. CRISPR-on has been proved to be effective in human and mouse. We expect to conduct similar work in our bio-factory, ''Escherichia coli''. | ||
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Our plan is to fuse alpha factors with dCas9 protein, which has previously been proved to be a successful method to create blue-light-induced transcription factors (Camsund et al., 2011). Our design is shown below. | Our plan is to fuse alpha factors with dCas9 protein, which has previously been proved to be a successful method to create blue-light-induced transcription factors (Camsund et al., 2011). Our design is shown below. | ||
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[[File:CRISPR-on.png|thumb|600px]] | [[File:CRISPR-on.png|thumb|600px]] | ||
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However, to integrate CRISPRi and CRISPR-on would never be an easy task, since sgRNA for CRISPR-on is supposed to target upstream of promoter, rendering it necessary to incorporate logical switches. | However, to integrate CRISPRi and CRISPR-on would never be an easy task, since sgRNA for CRISPR-on is supposed to target upstream of promoter, rendering it necessary to incorporate logical switches. | ||
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=Smaller Light Sensor= | =Smaller Light Sensor= | ||
Revision as of 03:29, 28 September 2013
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