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Notebook: April
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Thursday, April 14
- Today we had our first meeting with a grad student- just bouncing some ideas off of him for feasibility. There is no RNAi in prokaryotes! Some ideas included a small molecule inhibitor / morpholino oligomers.
- The first CRISPR information was found and was noted as something to follow up on in further meetings.
Thursday, April 21
- We discussed CRISPR more exensively- we are beginning to latch onto the idea as a fully formed project. Building on earlier discussions, the goal would be to target an antibiotic resistance gene (NDM-1) to ameliorate that resistance completely.
Thursday, April 28
- Ethan presented an outline of CRISPR and the possible project to the group for consideration. Some questions that came up:
- What is the advantage of using this entire process? Is not it kind of roundabout?
- The structure of CRISPR lends itself to a modular platform- the degree to which the pathway is active can be shown to be dependent on the number of spacers which are easily controlled.
- How will we motivate bacteria to take up potentially deleterious genetic material? What about an F-factor plasmid?
- Trojan Horse D- maintains growth advantage
- What has been done, what is the advantage of this approach?
- Practical point of view: two steps that can be done in parallel
- CRISPR design for different sequences
- Downregulate NDM1 protein with plasmid
- Can we use phages to introduce a CRISPR array?
- High throughput methods & random mutations, screen for them
- Could we use CRISPR to downregulate a repressor of something?
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