Analysis of the Role of NuMA (TDIF) in DNA repair Through Next Generation Sequencing Techniques [PolII ChIP-seq RPE1]

Purpose: To assess NuMA (TDIF)’s role in facilitating PolII pausing Methods:Pol2 ChIP-Seq was carried out in each pairwise combination of untreated/hydrogen peroxide treated and no doxycyclin (NuMA (TDIF) WT)/doxycyclin treated (NuMA (TDIF) KD) RPE1 cells in duplicate. The data was mapped using bwa mem and subsequently used to generate bigwigs showing fold-change relative to inputs using MACS2 and various meta-feature profiles using the CGAT suite of bioinformatics tools Results: The data revealed increased occupancy of PolII at promoter regions and increased pausing ratios for genes in H2O2- cells after NuMA (TDIF) KD relative to controls, but decreased occupancy in NuMA (TDIF) KD in H2O2+ cells compared to controls. Conclusions: Our study indicates that NuMA (TDIF) plays a role in PolII pause-release. Assessment of PolII binding within the genome through ChIP-seq in RPE1 cells under untreated/hydrogen peroxide treated conditions and no doxycyclin (NuMA (TDIF) WT)/doxycyclin treated (NuMA (TDIF) KD) conditions in duplicate MH = Untreated/H2O2-, PH = H2O2 treated, ND = No doxycyclin/WT with respect to NuMA (TDIF), PD = post-doxycyclin/NuMA (TDIF) KD, IN = Input, IG = IgG, Pol2 = RNA polymerase II