Team:UCSF

From 2013.igem.org

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The human race is growing fast. The world population is projected to top 10 billion by 2050. One response to the food supply needs of an increasing world population is creating better crop yield through crop survival. Pathogens and pests can cause catastrophic crop loss. Currently there is no widely used synthetic biological mechanism to help plants fight off bacterial pathogens and pests, so the main way large-scale growers protect crops is by using pesticides. We have engineered two possible solutions to address this problem.
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In microbial communities, bacterial populations are commonly controlled using indiscriminate, broad range antibiotics. There are few ways to target specific strains effectively without disrupting the entire microbiome and local environment. The goal of our project is to take advantage of a natural horizontal gene transfer mechanism in bacteria to precisely affect gene expression in selected strains.  We combine bacterial conjugation with CRISPRi, an RNAi-like repression system developed from bacteria, to achieve targeted control of gene expression within a complex microbial community. One possible application is to selectively repress pathogenic genes in a microbiome, leaving the community makeup unaffected. In addition, we use CRISPRi to lay the groundwork for transferring large circuits that enable complex functionality and decision-making in cells.
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In our first project, we have incorporated catalytically dead CAS9 (dCAS9) in conjunction with synthetic gRNAs complementary to pathogenic bacterial genes into a plasmid that naturally conjugates into neighboring bacterial cells. The specificity of the gRNAs will potentially allow us to target specific populations of bacterial pathogens in soil while leaving positive bacterial populations unharmed.
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Our second project will provide plants with a sugar-mediated synthetic circuit that boosts a plant’s natural immune system and pesticide-producing capabilities in response to different levels of sugar a plant naturally produces in response to pests. The repression capabilities of dCAS9 will be used to regulate the synthetic circuit promoters that sense high or low sugar levels, reducing metabolic strain.
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By using these two synthetic biology solutions, we hope to be able to positively impact crop production while minimizing chemical inputs, in a way that will benefit the world’s food supply and environment.
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Revision as of 18:01, 30 August 2013

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ABSTRACT


In microbial communities, bacterial populations are commonly controlled using indiscriminate, broad range antibiotics. There are few ways to target specific strains effectively without disrupting the entire microbiome and local environment. The goal of our project is to take advantage of a natural horizontal gene transfer mechanism in bacteria to precisely affect gene expression in selected strains. We combine bacterial conjugation with CRISPRi, an RNAi-like repression system developed from bacteria, to achieve targeted control of gene expression within a complex microbial community. One possible application is to selectively repress pathogenic genes in a microbiome, leaving the community makeup unaffected. In addition, we use CRISPRi to lay the groundwork for transferring large circuits that enable complex functionality and decision-making in cells.



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