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Team:UCSF/Notebook
From 2009.igem.org
HL60 Group
- Goals: Our goals for this project are to screen 30 various different GPCR's (G couple protein receptors)to determine which receptors mediate chemotaxis using the high through put transwell assay. After first phase screening using 5-6 day differentiated HL-60 cell transfected with known receptors is completed, we move on to the secondary stage of screening with viable chemotatic receptors. During the second stage of screening we fused "Actin Modulators", "PIP3 Modulators", and "GEF Activators" in hopes of an accelerated chemotatic response.
- Engineering Navigation:
- sending cells to new targets - GPCR Screening
- tuning receptor sensitivity - Forced localization of effector proteins to GPCRs
- Payload:
- can we make our cells carry stuff? - attach beads to cells
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- Cathy Liu's notebook: I was one of the main players on team HL-60 who conducted the transwells for the GPCR screen. This included transfections and analysis of data. I was Aynur’s student and Ryan’s and Eric’s buddy. And what a beautiful team we made!
- Eric Wong's notebook: I worked on cloning, PCR, Gel purification and extraction, and was apart of the GPCR screening team.
- Jackie Tam's notebook: I was responsible for cloning, microscopy, and video analysis.
- Ryan Liang's notebook:I cloned parts to be used in the GPCR screens, transfected HL-60 cells with various different GPCRs, ran transwell assays, and analyzed/compared transwell assay results.
- Katja Kolar's notebook: I did microscopy with wt and hM4D-transfected HL-60 cells, experiments for the "Payload" part of the project, and cloned HL-60 team constructs.
- Hansi Liu's notebook:
Dictyostelium Group
- Goals:
- Engineering brakes and accelerators:
- modulating PIP3 polarity by building synthetic protein based feedback loops
- workflow:
- 1. PCR parts: catalytic and localization domains
- 2. Create combinations of localization domains and catalytic domains
- 3. Generate Dictyostelium strains expressing feedback elements
- 4. Measure motility parameters (speed, directionality)
- 3. Analyze data. Create histograms and compare if our plasmid had any effect to dicty.Do they cause them to move faster or slower?
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- Allen Cai's notebook: I worked mostly on the clonning part of the project. I ligated a lot of the localization and catalytic domains together. I also took care so some dicty strains and made some movies of those dicty strains.
- Alex Smith's notebook:
- Edna Miao's notebook: I transformed new constructs into Dicty, analyzed data, and worked with wildtype and PTEN null dicty.
- Ethan Chan's notebook: Constructed Dicty cells with new parts, analyzed data, and worked with wildtype and PTEN null dicty cells.
- Ryan Quan's notebook:I worked on team dicty and taught the students some basic cloning techniques but mainly learned many new things along with them.
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