Question

Write no more than necessary to answer the question. Pictures are encouraged if they would be helpful. For all questions, you may assume that high quality reagents are available and all relevant DNA sequences have been cloned. Money is no object, but experiments should be focused on answering the question. 

You are a postdoc at the Jedi Institute for Advanced Studies. Your project is focused on the cellular pathway regulating sensitivity to the Force. Pedigree analysis suggests that Force sensitivity runs in families and is inherited in a Mendelian dominant fashion. Classical genetic and mapping studies performed during the Old Republic localized the gene for Force sensitivity to the very end of the P arm of chromosome 3 (locus C3p0).  Using bioinformatics analysis, a collaborator has discovered a polymorphism in one of the open reading frames in this region (provisionally named FORCE) that correlates strongly with sensitivity to the Force. 

You hypothesize that the wild type FORCE protein represses genes that are involved in the signaling pathway that responds to the Force. The mutation results in the abnormal upregulation of these genes, and thus increased sensitivity to stimulation of the pathway. 

****Describe an experiment to determine which genes are functionally regulated by FORCE *****

Answer 1

1. Over express the force repressor and observe the expression of gene by q real time PCR

Help to identification of gene which are not express.

2. Knockout the fore and observe the expression by q real time PCR.

Help to identification of gene which activated after knock out

3. Confirm the result by western blot.

4.ChIP-sequencing help to understand actual gene which repress by FORCE  

This four experimental help to determine gene which regulated by FORCE.

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