Difference between revisions of "Team:Tufts"
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The CRISPR-Cas9 system is a revolutionary new tool in molecular biology, allowing for double-stranded DNA breaks to be made almost anywhere in the genome. This technology makes it possible to alter targeted genes in an organism with the inclusion of a guide RNA that specifies where the Cas9 endonuclease cuts. CRISPR/Cas9 can be repurposed as a therapeutic gene-editing platform to treat genetic disorders, by replacing a mutated gene with a functional version. | The CRISPR-Cas9 system is a revolutionary new tool in molecular biology, allowing for double-stranded DNA breaks to be made almost anywhere in the genome. This technology makes it possible to alter targeted genes in an organism with the inclusion of a guide RNA that specifies where the Cas9 endonuclease cuts. CRISPR/Cas9 can be repurposed as a therapeutic gene-editing platform to treat genetic disorders, by replacing a mutated gene with a functional version. | ||
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Currently, we have received the plasmids for the atoxic TcdB and the Cas9 protein and have begun culturing them in E. coli. We plan to express the modified toxin in B. megaterium and then deliver the fusion protein to human epithelial cells expressing GFP. We have designed a proof of concept experiment to test for effective delivery of the Cas9 protein in which we will attempt to remove the GFP coding region in epithelial cells via this proposed CRISPR/Cas9 system. We will measure the fluorescence of the cells after TcdB/Cas9 fusion protein has been delivered, hoping to see a decrease in fluorescence as a result of the GFP coding region being removed by Cas9. | Currently, we have received the plasmids for the atoxic TcdB and the Cas9 protein and have begun culturing them in E. coli. We plan to express the modified toxin in B. megaterium and then deliver the fusion protein to human epithelial cells expressing GFP. We have designed a proof of concept experiment to test for effective delivery of the Cas9 protein in which we will attempt to remove the GFP coding region in epithelial cells via this proposed CRISPR/Cas9 system. We will measure the fluorescence of the cells after TcdB/Cas9 fusion protein has been delivered, hoping to see a decrease in fluorescence as a result of the GFP coding region being removed by Cas9. | ||
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<h2> Welcome to iGEM 2015! </h2> | <h2> Welcome to iGEM 2015! </h2> | ||
Revision as of 02:51, 10 September 2015
