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Team:UCSF/Project/Accomplish1
From 2013.igem.org
We have shown that CRISPRi can be introduced to cells through bacterial conjugation. This is the first time that this has been shown and provides the opportunity for many future applications. We have shown that after being introduced by conjugation, our CRISPRi system is functional and capable of repressing a targeted gene (RFP). We have created a standard for creating "knock-in" cassettes and inserting DNA into the E. coli genome. Using a combination of PCR sewing and lambda red recombination, we inserted two fluorescent proteins into the E. coli genome to mark our conjugation strains. Our protocol was tested several times and has proven to be very effective. Write-up for our Standard We have created a set of materials that can be used to introduce synthetic biology to the public. These materials were produced as a part of our outreach and human practices projects and are available for download on our Materials page. We have further characterized the pTET and pLAC promoters that are widely used in the iGEM registry and placed this data on the parts registry pages.
