Suppose your research team found a brand new gene that may be involved in the early stages of embryonic development. In studying this gene, your team manages to identify the promoter region and one enhancer region that is 1,000 kilobases upstream of the promoter region. To study this gene’s expression during embryonic development, would you choose to fuse a reporter gene, like GFP, with the promoter region or the enhancer of your gene of interest? Why would you choose that region for your studies?
Answer: Both promoter and enhancer regions must be taken.
Explanation: The endogenous expression of a gene is dependent upon all of its cis-acting elements (promoter, enhancers, and silencers) as well as the presence of its respective trans-acting factors. The promoter is bound by RNA polymerase. The enhancer is a cis-element bound by an activator and induces gene expression. Silencer is bound by a repressor and represses gene expression.
The endogenous spatial and temporal expression pattern is governed by the collective outcome of the interaction between all the cis elements and trans-factors. Each interaction is sensitive to specific developmental or exogenous signal. So, if we want to analyze the endogenous expression pattern, we must include all the known/available cis-elements in the reporter construct.
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