DNA vector-based RNAi approach for stable depletion of poly(ADP-ribose) polymerase-1
- Laboratory for Skin Cancer Research, CHUL Research Center (CHUQ), Faculty of Medicine, Laval University, Sainte-Foy, Que., G1V 4G2 (Canada)
RNA-mediated interference (RNAi) is a powerful technique that is now being used in mammalian cells to specifically silence a gene. Some recent studies have used this technique to achieve variable extent of depletion of a nuclear enzyme poly(ADP-ribose) polymerase-1 (PARP-1). These studies reported either transient silencing of PARP-1 using double-stranded RNA or stable silencing of PARP-1 with a DNA vector which was introduced by a viral delivery system. In contrast, here we report that a simple RNAi approach which utilizes a pBS-U6-based DNA vector containing strategically selected PARP-1 targeting sequence, introduced in the cells by conventional CaPO{sub 4} protocol, can be used to achieve stable and specific silencing of PARP-1 in different types of cells. We also provide a detailed strategy for selection and cloning of PARP-1-targeting sequences for the DNA vector, and demonstrate that this technique does not affect expression of its closest functional homolog PARP-2.
- OSTI ID:
- 20709204
- Journal Information:
- Biochemical and Biophysical Research Communications, Vol. 331, Issue 1; Other Information: DOI: 10.1016/j.bbrc.2005.03.135; PII: S0006-291X(05)00626-1; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
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