E6AP gene suppression and characterization with in vitro selected hammerhead ribozymes.

Abstract:

:E6AP was originally identified as the ubiquitin-protein ligase involved in human papillomavirus (HPV) E6-mediated p53 degradation and has since been shown to act as an E3 ubiquitin-protein ligase in the ubiquitination of several other protein substrates. To further define E6AP function at the molecular and cellular levels, a ribozyme-based gene inactivation approach was adopted. A library of hammerhead ribozymes, with randomized arm sequences, was used to screen active molecules along the entire E6AP transcript for ribozyme-cleavable sites. Ligation-anchored PCR was adapted to detect cleavage products, and ribozymes designed to the selected sites were characterized both in vitro and in vivo. Ribozyme-mediated reduction in E6AP expression was found to enhance the apoptotic response of HeLa cells to mitomycin C-induced DNA damage. These findings suggest that E6AP has potential as a drug target, as its suppression can potentiate apoptosis in HPV-positive cells treated with a cytotoxic drug.

journal_name

Cancer Gene Ther

journal_title

Cancer gene therapy

authors

Kim Y,Cairns MJ,Marouga R,Sun LQ

doi

10.1038/sj.cgt.7700623

subject

Has Abstract

pub_date

2003-09-01 00:00:00

pages

707-16

issue

9

eissn

0929-1903

issn

1476-5500

pii

7700623

journal_volume

10

pub_type

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