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Team:WHU-China/Project
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=== Part 2 === | === Part 2 === | ||
| - | + | In this part, we are going to deal with the targeting system. We fuse dCas9 protein with another two proteins, cI repressors and Gal11. Both of them is cloned from Escherichia coli and have been tested for their interaction with other proteins we use in our second system. | |
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=== The Experiments === | === The Experiments === | ||
Revision as of 09:01, 9 August 2013
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Contents |
Overall project
Prokaryotic type II CRISPR-Cas systems are quite powerful compared with RNAi or TALEN and they can be adapted to enable targeted genome modifications across a range of organisms, both prokaryotes and eukaryotes. In spite of the rapid explosion of research concerning with Cas system, their applications are quite limited, especially considering the gene activation.
In order to further explore the detailed functions of Cas system, we design a novel platform on which a multistage gene regulation can be achieved precisely as expected. In our project, we engineer two fusion systems associating with each other via protein-protein interactions. First, a nuclease-null Cas9 protein along with various sgRNA is fused with either Gal11 or cI repressors. They can bind specificly to its targets and interact with the second system. In the second part, alpha factor, a key element of prokaryotic transcription complex, is coexpressed with another four proteins, including Gal4-1,Gal4-2,cI and VP16. Their interactions is powerful enough to recruite more elements to form the transcription complex. As a consequence, downstream reporter gene can be either activated or silenced according to the specific forms of sgRNA as described above.
Project Details
In our project, there are two systems, one of which is applied to target different parts of a single promoter. Another part is designed to recruite disparate elements to assemble the transcription complex.
Part 2
In this part, we are going to deal with the targeting system. We fuse dCas9 protein with another two proteins, cI repressors and Gal11. Both of them is cloned from Escherichia coli and have been tested for their interaction with other proteins we use in our second system.
