Optimization of ChIP protocol for investigation of canonical Wnt target genes expression in new born heart

  • O. O. Piven
DOI: https://doi.org/10.7124/FEEO.v22.943

Abstract

Aim. In present work we have focused on chromatinimmunoprecipitation (ChIP) protocol with next qPCR optimization for analysis of canonical Wnt target genes regulation by b‑catenin and plakoglobin in new born heart. Methods. In study we have used the new born hearts tissue at the age of 3 days after birth (P3). We have analyzed hearts of control mice and mice with heterozygous knockout of β-catenin. We have proceeded ChIP protocol based on standard protocols in our modifications. ChIP results validated with qPCR. Results. We have adopted ChIP protocol for investigation of signalling function of β-catenin and plakoglobin as well as for identification of β-catenin and plakoglobin specific binding genomic region in new born heart. Conclusions. We have shown that plakoglobin is able to compensate the signaling function of β-catenin and binds with similar genomic region under β-catenin haploinsufficiency conditions.

Keywords: β-catenin, plakoglobin, Wnt signaling, gene expression, heart, chromatinimmunoprecipitation, ChIP.

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