a) FLC gene expression is suppressed following vernalization. Describe two epigenetic histone modifications that lead to FLC gene suppression.

b) These histone modifications on the FLC gene were originally detected by chromatin immunoprecipitation (ChIP). Using antibodies specific for each of the modified histones (part a) in a ChIP experiment, would the amount of detected FLC DNA increase or decrease during the vernalization period? Explain your answers.

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Keith LeannonLv2

19 Mar 2019

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In your research you are analyzing the changes in liver gene expression in a mouse model of non-alcoholic fatty liver disease. You previously performed microarray analysis on livers of mice given a high-fat diet verses mice on regular diet and found a number of genes that were abnormally expressed in the high-fat diet mice. You perform quantitative RT-PCR on some of these genes and receive the following Ct values:

high-fat diet: Gapdh-10, Cyp8b1-27, Cyp17a1-25, Asns-21, Past1-23.

Regular diet: Gapdh-11, Cyp8b1-25, Cyp17a1-21, Asns-26, Past1-30.

Question a: For each of the genes, complete calculations using the delta-delta Ct method to determine the relative fold change in gene expression for the mice with the high-fat diet. (hint use Gapdh as the reference gene and SHOW YOUW WORK).

Question b: Which of the four genes are up-regulated and which of the four genes are down-regulated in mice with high-fat diets?

Question c: You want to determine exactly where in the liver these genes are normally expressed. Pick one of the genes above and design an experiment to perform in livers of high-fat diet mice and regular diet mice (as a control) to determine the location of the gene expression in the liver. Describe the results you expect to see from this experiment in the two different mouse livers.

Question d: You hypothesize that the change in expression of the gene you picked in part C. Above is a result of differences in epigenetic marks. Briefly describe what changes in epigenetic marks at this gene's locus are expected to occur as a result of the high-fat diet. In your answer address both DNA methylation and histone modifications.