Faculty Advisor

Adams, David S.

Abstract

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.

Publisher

Worcester Polytechnic Institute

Date Accepted

January 2004

Major

Biology and Biotechnology

Project Type

Major Qualifying Project

Accessibility

Restricted-WPI community only

Advisor Department

Biology and Biotechnology

Advisor Program

Biology and Biotechnology

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