Gene Repair: Depletion of PADPRP by Antisense RNA Expression
Abstract
To define the role of poly(ADP-ribose) polymerase (PADPRP) in DNA repair, we have developed cell lines from human HeLa cells, that express abundant PADPRP antisense transcripts under the control of MMTV promoter. The depletion of endogenous PADPRP by induced antisense RNA expression was established by: (1) a progressive synthesis of antisense transcripts in cells as assessed by Northern analysis; (2) an 80% decrease in activity of the enzyme; and (3) a greater than 90% reduction in the cellular content of PADPRP protein, as demonstrated by both immunoblotting and immunohistochemical analysis in intact cells. In cells expressing antisense RNA of PADPRP the ability to repair MMS-induced DNA single-strand break is totally inhibited at 20 minutes, partially recovered after 45 minutes, and fully recovered after 90 minutes indicating that the low amount of PADPRP enzyme delayed the DNA single-strand break repair.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 13, 1993
- Accession Number
- ADP008775
Entities
People
- Mark Smulson
- Ruchuang Ding
- Tina Stevnsner
- Vilhelm Bohr
Organizations
- Georgetown University