Protocol

Poly(ADP-ribose) Polymerase

Volume 780 of the series Methods in Molecular Biology pp 191-208

Date:

PARP1 Genomics: Chromatin Immunoprecipitation Approach Using Anti-PARP1 Antibody (ChIP and ChIP-seq)

  • Niraj LodhiAffiliated withEpigenetics and Progenitor Cells Program, Fox Chase Cancer Center
  • , Alexei V. TulinAffiliated withEpigenetics and Progenitor Cells Program, Cancer Biology Program, Fox Chase Cancer Center

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Abstract

Poly(ADP-ribose) polymerase1 (PARP1) is a global regulator of different cellular mechanisms, ranging from DNA damage repair to control of gene expression. Since PARP1 protein and pADPr have been shown to persist in chromatin through cell cycle, they may both act as epigenetic markers. However, it is not known how many loci are occupied by PARP1 protein during mitosis genome-wide. To reveal the genome-wide PARP1 binding sites, we used the ChIP-seq approach, an emerging technique to study genome-wide PARP1 protein interaction with chromatin. Here, we describe how to perform ChIP-seq in the context of PARP1 binding sites identification in chromatin, using human embryonic kidney cell lines.

Key words

PARP1 Genomics Human embryonic cells Mitotic arrest ChIP ChIP-seq