1. State the problem to be investigated and explain why the topic is important or of interest. 2. Summarize the current knowledge related to the project and highlight the relevant data (Use data to explain the relevance of multiple myeloma among men and women) 3. Provide the rationale for the proposed research –> explain why the specific research questions are important (because current treatments are associated with poor survival rates among patients) 4. The hypothesis should be clearly stated, focused and understandable – should be feasible and testable – should be supported by the background information 5. Provide expectations or explain how the results obtained will be used to test the hypothesis 6. The aims should be designed to test the hypothesis 7. Enough explanation of what will be done in each aim 8. State the expected outcomes 9. Explain the experimental approaches use these papers; they are really helpful 1. Neri P, Ren L, Azab AK, Brentnall M, Gratton K, Klimowicz AC, et al. Integrin β7-mediated regulation of multiple myeloma cell adhesion, migration, and invasion. Blood. 2011 Jan 1. 2. Multiple myelomas: biology of the disease Anuj Mahindraa, *, Teru Hideshimab, Kenneth C. Andersonb Extra information that you need to complete this assignment: – We knock CRBN with CRISPER-Cas9 – We need to be efficient in the knockdown to see the effect – We are only focusing on the IMiDs –> lenalidomide (selective for multiple myeloma) – we are using a comprehensive 12 myeloma cell lines used to represent different sub-groups of myeloma (in terms of genetic alterations, translocation, and deletions) – another methodology is transfection (have a control = empty vector) and the transfected – To look for the efficiency of transfection; we will use quantitative PCR(look for the mRNA) and (Western Blot=prtotein expression) – When doing western blot we look for Cereblon and the IKZF expressions – AIM 2: sometimes it is not really CRBN that makes the cell resistant but a specific region in that gene that’s why we are examining exon 10 (because it is overexpressed) – myeloid cells express more Cereblon – RNAseq data from patients is already available for the analysis Main study reason: evaluating the drug mentioned above and understanding its drug-resistant mechanism – The hypothesis is that cereblon is important and may be used as a biomarker (correlation between cereblon expression and respone) – we are hoping to use cereblon as a biomarker of response/resistant in myeloma patients.

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