Team:SJTU-BioX-Shanghai/Prospect

From 2013.igem.org

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(CRISPRi-on)
(CRISPRi-on)
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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''.
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]]
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[[File:CRISPR-on.png|thumb|700px]]
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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.
=Smaller Light Sensor=
=Smaller Light Sensor=

Revision as of 03:27, 28 September 2013

CRISPRi-on

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. 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. 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.

CRISPR-on.png

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.

Smaller Light Sensor

Absolutely Automatic