Interesting findings by ChIP-Seq and DNA-Seq analysis using Strand NGS

The year 2014 was a great year with exciting features and enhancements updated in Strand NGS. We take this opportunity to thank our clientele and well-wishers for their support and feedback. We wish you all a happy and prosperous new year and look forward to a more fruitful engagement in 2015.
In this blog on Interesting findings by ChIP-Seq and DNA-Seq analysis using Strand NGS, we present two recent publications that analyse ChIP-Seq and DNA-Seq data using Strand NGS.
1. A Comprehensive Profile of ChIP-Seq-Based PU.1/Spi1 Target Genes in Microglia by Satoh et al.
2. Localised Dominant Dystrophic Epidermolysis Bullosa with a Novel de Novo Mutation in COL7A1 Diagnosed by Next-generation sequencing by Nagai et al.

The paper by Satoh et al. investigates the biological role of the transcription factor PU.1 in regulation of microglial functions. Though PU.1 plays vital role in microgliogenesis, the comprehensive profile of PU.1/Spi1 target genes in microglia is unknown. In this paper, Strand NGS was used to analyse SRP036026 ChIP-Seq data set and identify the role of PU.1/ Spi1 in microglial gene regulation. Using Strand NGS, around 5,264 ChIP-Seq-based Spi1 target protein coding genes (Spi1, Irf8, Runx1, Csf1r, Csf1, Il34, Aif1 (Iba1), Cx3cr1, Trem2, and Tyrobp) were identified in BV2 mouse microglial cells. Motif analysis by GADEM revealed the PU-box consensus sequences (5’-GAGGAA-3’) were located on 80.3% of the peaks detected by MACS. By downstream pathway analysis, the ChIP-Seq-based Spi1 target genes were found to show significant relationship with diverse pathways essential for normal function of monocytes/ macrophages (like endocytosis, phagocytosis, lysosomal degradation). Hence PU.1/Spi1 was found to have an important role in microglial gene regulation and any aberrant regulation of these target genes would contribute to neurodegenerative diseases by activated microglia accumulation.

The second paper by Nagai et al. is a case study of a 10-month-old female infant with local Dominant Dystrophic Epidermolysis Bullosa. A targeted next generation sequencing was performed with the proband’s peripheral blood for 16 genes associated with Dystrophic Epidermolysis Bullosa. The sequenced data was aligned and analysed using the DNA variant analysis workflow in Strand NGS (formerly Avadis NGS). A heterozygous single nucleotide variation on chr.3: g.48616827C>T (negative strand) that corresponds to a missense mutation of p.Gly1761Asp in the triple helix domain of COL7A1 was detected. This de novo high confidence mutation was also confirmed by Sanger sequencing. This mutation was detected only in the proband and was not found in the parents, in 100 healthy Japanese alleles as well as in dbSNP.