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Exploration of the Role of Negative Elongation Factor on Tat Transactivation using siRNAs

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Transcriptional regulation at the elongation step has recently been the focus of intense research. Several proteins have been identified which possess either positive or negative effects on this process; negative elongation factor (NELF) is one such protein. The role of NELF on HIV-1 transcriptional elongation has yet to be fully characterized. Using the newly developed RNAi technology, we attempted to significantly reduce cellular expression of an essential subunit of NELF, NELF-E, in MAGI cells to help elucidate its role in Tat-transactivation. Results from RT-PCR and immunoblotting in conjunction with data from a beta-galactosidase reporter gene assay monitored knockdown and the effects on Tat-transactivation. Successful knockdown was achieved for positive control Spt5, but not for NELF-E. Sequencing of selected regions of the NELF-E gene in MAGI cells indicated no mismatches with the published NELF-E sequence upon which the siRNAs were based.

  • This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.
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  • 04C005M
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  • 2004
Date created
  • 2004-01-01
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