Team:UCSF/Project/Background2
From 2013.igem.org
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<p2>At the beginning of this summer, we asked ourselves a question: "What could we introduce to a microbiome which would allow targeting and eventual gene expression changes in a specific bacteria?" The difficulty faced with this situation is in:</p2> | <p2>At the beginning of this summer, we asked ourselves a question: "What could we introduce to a microbiome which would allow targeting and eventual gene expression changes in a specific bacteria?" The difficulty faced with this situation is in:</p2> | ||
<p2> 1. Introduce a targeting system into a defined mixture of bacteria such that you can select and introduce manipulations without negatively affecting other bacteria. </p2> | <p2> 1. Introduce a targeting system into a defined mixture of bacteria such that you can select and introduce manipulations without negatively affecting other bacteria. </p2> | ||
<p2> 2. Creating easy to transfer pathways or circuits that can produce a multitude of outcomes (killing, repressing, upregulating). </p2> | <p2> 2. Creating easy to transfer pathways or circuits that can produce a multitude of outcomes (killing, repressing, upregulating). </p2> | ||
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<h3>1. Introducing CRISPRi to a bacterial community</h3> | <h3>1. Introducing CRISPRi to a bacterial community</h3> | ||
<p2>To selectively target and eliminate harmful bacteria, we are utilizing the CRISPRi system, a tool repurposed from a natural adaptive immunity system in bacteria (see diagram below). This tool is comprised of a catalytically dead Cas9 (dCas9) protein that complexes with guide RNAs (gRNA) complementary to the target bacteria’s DNA sequence. This complex binds to DNA complementary to the gRNA and prevents transcription, therefore repressing gene expression.</p2> | <p2>To selectively target and eliminate harmful bacteria, we are utilizing the CRISPRi system, a tool repurposed from a natural adaptive immunity system in bacteria (see diagram below). This tool is comprised of a catalytically dead Cas9 (dCas9) protein that complexes with guide RNAs (gRNA) complementary to the target bacteria’s DNA sequence. This complex binds to DNA complementary to the gRNA and prevents transcription, therefore repressing gene expression.</p2> | ||
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<img style="height:400px"; padding:0;" src="https://static.igem.org/mediawiki/2013/5/56/CRISPRi_Background_UCSF.png"> </div> | <img style="height:400px"; padding:0;" src="https://static.igem.org/mediawiki/2013/5/56/CRISPRi_Background_UCSF.png"> </div> | ||
Revision as of 08:23, 28 October 2013
Operation CRISPR: Deploying precision guided tools to target unique species in a complex microbiome
1. Introducing CRISPRi to a bacterial community
WHY USE CRISPRi?
1. CRISPRi utilizes gRNAs which are highly specific and customizable.
2. In principle it could be used to take advantage of unique DNA sequences to target specific bacterial species.