Project “Grant” Proposal - Do not plagiarize - How should you structure your project proposal assignment? Refer to this NIH grant writing suggestions website: http://www.biosciencewriters.com/NIH-GrantApplications-The-Anatomy-of-a-Specific-Aims-Page.aspx - Make sure you ask questions if anything is unclear APA format 3 pages double; max 4 pages spaced NOT including reference page Use the links provided Once I accept your bid I will post the protocol for the invasion and colony formation experiment ( MAKE SURE YOU READ THE PROTOCOLS TO UNDERSTAND HOW THE ASSAYS WORK) Part 1: The gene that will be used to carry out the experiments is Sh DCBL2 (RNAi) and the non-targeting control which is an scramble shRNA ( sh= short hairpin) → PANC-1 CELLS This gene is an oncogene and we want to downregulate it on the experiment shDCBLD2 Turning off oncogene DCBLD2 is prognostic, high expression is unfavorable in pancreatic cancer. shRNA disables gene expression 1) To summarize and critically evaluate the literature to establish current knowledge relating to the gene/protein whose expression is being upregulated by CRISPR activation in the pancreatic cancer cells that you are studying. 2) To demonstrate the GAPS that exist in the current knowledge about your subject and what your solutions is to solve them. In this section, you should include a hypothesis regarding your approach. 3) To come up with two specific aims for your (end of semester) independent project and briefly describe how they work to test your hypothesis. a. Aim 1: should be a transwell invasion assay b. Aim 2: colony formation assay we will be using a dox-inducible (drug) on the PANC-1 shRNA cell lines (DCBL2 and scrambled shRNA control cell line) to determine the impacts of invasion and colony formation Part 2: 1) Experimental Plan: List each specific aim separately and describe in detail how you will carry out an experiment(s) to achieve each aim. Topic: You will be given a CRISPR-activation or shRNA knockdown pancreatic cancer cell line to study for this project. As a reminder, these cell lines have been modified to increase or decrease the expression of the genes of interest. This is because previous studies determined that each of these genes is underexpressed or overexpressed in the progression from early to late stage pancreatic cancer in human tumor tissue samples. Thus, downregulation/ upregulation of these genes may be correlated to cancer progression. However, we currently do not know the precise role of these genes in the development or progression of pancreatic cancer. Backgound Research: PubMed searches using your gene name and “cancer” will return articles that will give you an idea of what is known about the role of your gene and cancer. Focus on the introduction of these articles and any review articles that you can find. You can also type in specific types of cancer to determine what role it plays in particular types of cancer, including pancreatic, if any. Here are a couple of articles I have already found: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4151226/ Potential therapeutic target http://mcr.aacrjournals.org/content/6/2/222.long Epigenetic Down-Regulation and Suppressive Role of DCBLD2 in Gastric Cancer Cell Proliferation and Invasion Sources: At least 3 primary research articles must be referenced (APA-style) in your proposal; inclusion of additional resources are up to you. Length: 3 pages, double spaced; maximum 4 pages. How should you structure your project proposal assignment? Refer to this NIH grant writing suggestions website: http://www.biosciencewriters.com/NIH-Grant-Applications-TheAnatomy-of-a-Specific-Aims-Page.aspx