Questions for Lecture 26: Genetic screen for HIV-infectivity 1) To examine at what step Rab6 is required for HIV infection, Rab6 knockdown cells were infected with either (1) normal HIV (which binds to CD4) or (2) HIV with a virion surface that binds to a different human cell receptor. Only normal HIV had reduced infectivity, while HIV entry via a different receptor produced a normal infection level. In the same assay, what result would you expect with the TNPO3 knockdown? 2) TNPO3 knockdown cells were tested for reverse transcriptase (RT) activity in the cytoplasm after infection. The RT activity was normal. What if the RT activity were, instead, reduced compare to the negative control (Luciferase knockdown). What step(s) are probably affected by TNPO3 knockdown? 3) You wish to reproduce the HIV-infectivity screen in your laboratory, and you have developed your own assay. When you test the assay with controls, you get the following results: siRNA gene target for knockdown Percent infected (+/- standard error)
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