Global transcriptomic analysis of Drosophila Hinfp mutant and wild-type gut to understand their role in genome stability

Drosophila hinfp mutant is lethal. They are unble to ecolose from the pupal case and died. To investigate the molecular function of Hinfp in Drosophila gut, we isolated RNA from mutant and wild type adult pharate gut and performed RNA-seq analysis. The comparison of mutant and control samples revealed significantly changed expression of over more than thousand Drosophila genes. Among the down regulated genes were many Histone genes. Further validation of RNA-seq data of Histone genes divulged that His1 is significantly down-regulated lead to genome instability through de-repression of menay of the retrotransposon. We also rescued the all phenotypes by overexpression of UAS-Hinfp and UAS-His1 gene. Fifty midguts with malpighian tubules were dissected from 5-6 days old control and 8-9 days old mutants and rescue male adults pharate and adult cultured at 25°C. The genotypes of the flies were, Y/FRT19A; +/+; +/+, Y/Hinfp2; +/+; +/+ and Y/Hinfp2; +/+; tub-Gal4/UAS-Hinfp-HA, Y/Hinfp2; +/+; tub-Gal4/UAS-His1-HA. Total RNA was extracted by Tri- reagent (Sigma, USA) according to the instructions, followed by DNAas I (catalog no. 79254; Qiagen) and purified with RNA-easy kit (Qiagen (catalog no. 74104; Qiagen)). RNA was quantified at by absorbance at 260 nm using a ND-1000 spectrophotometer (Nanodrop Technology, USA) and quality was assessed using a Bioanalyzer 2100 (Agilent Technology, USA) from UMMS molecular Biology core. All RNA-seq procedures start from quality, library construction using ribo-depletion kit to sequencing were carried out according to Company protocol (BGI, Hong Kong). The pair-end reads of 100 bp were done on the BGISEQ-500 platform for the following data analysis study.