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briefly explain how to do it for each aim, and 5) predict your results and analysis.

You are awarded for $1M grant to study a “genetic disease in modern society”.

First you need to choose a genetic disease and convince the review committee why the disease is “modern”.

Second, you are required to find out a gene closely related to the disease you choose and do some basic cloning work.

Finally, you need to do some studies on the disease. You will have all necessary reagents, equipment, and other resources for the study.
However, you are required to present a research proposal before you start.

Here are some specific requirements about your proposal:

1. The cover page should contain the title (must 80 or less characters) of your proposal, five keywords, and your name and date.

2. Diagram page: this page should have an overall diagram/carton to
summarize/highlight your project (called graphic abstract).

3. The 3rd page should contain general information about the disease you choose (similar to Assignment 2. You can explain why it is “modern” here).

4. The 4th page should contain a summary about the gene related to disease (similar to Assignment 3 first part).

5. The 5th page should contain DNA sequence information.
You need to list the name of your gene, accession number, species, size, etc. You also need to “copy/paste” the full-length “CDs” sequence. If your sequence is longer than half of a page, please cut the middle part of your sequence and put “////////////” sign there so that it can fit 2/3 of the page.
You need to calculate the protein size of your gene.
In addition, you need to design a pair of PCR primers for the whole CD sequence (not including “stop” codon).
Please highlight the primer positions on your sequence and write down the primer sequences in a right orientation (5’ to 3’) and Tms.
The primer should also contain restriction enzyme linkers for the cloning steps.

6. The 6th page should contain description about the cloning process.
1) how you “converted” the DNA sequence from computer database into your test tube;
2)how “clone” the gene into an expression vector;
3) how amplify the DNA in bacterial system;
4) how to purify the DNA; and
5) how to demonstrate your plasmid DNA is right one with a correct sequence.

7. The 7th page: you need to describe
1) how to deliver your cDNA plasmid into one of your targeted cells/tissues;
2) how to detect the expression of your gene;
3) how to demonstrate your gene indeed expressed, and illustrate the cellular localization of the protein. You need to use at least 3-4 technologies you learned in class to address these issues.

8. The 8th-9th pages should contain your research proposal. The proposal has to be specific and address one of the aspects of molecular cell biology, such as membrane transport, membrane trafficking, ER stress, mitochondrial defect, cell cytoskeleton, cell cycle, cell death, signaling transduction, etc.

You are encouraged to use the techniques you learned from the class in the study. The proposal should contains
1) rational (what problem you want to address and why it is important), 2) hypothesis: generally one sentence,
3) two specific aims,
4) briefly explain how to do it for each aim, and 5) predict your results and analysis.

9. Budget page (1 page): please list the dollar amount that you need for the study such as equipment, reagents, and fee for the facility, etc.

10. Reference page: list the references you used in your proposal. The style is same as Assignment 2 or 3.
11. The proposal should be typed, single space, 12-point font size, and 1-in marge around.

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