Exploration of RNA-Guided Recombinase Target Sites for Hyperactivated Recombinase Beta in Bovine Genome

Abstract

The present study was conducted to determine the effect of different cooling systems; Fan Fogger (FF) and Fan Pad (FP) on micro environment of poultry house, thermal comfort, welfare, egg production and egg quality parameters of laying hens. This experiment was conducted on 210, White Leghorn laying pullets (32 weeks old) during hot-dry summer months (May - July) under deep litter system of housing. The FP and FF cooling systems significantly dropped the mean shed temperature and increased the relative humidity. Thus, better THI resulted in increase in egg production by 4.66 % and 3.32 % under FP and FF systems over the control group. However, specific gravity, H.U, egg shell thickness, yolk index and yolk color were not significantly influenced by cooling treatments. Significantly lower levels of antioxidant enzymes viz. LPO, Catalase, G6PD, GPx and SOD was registered in cooling groups. Both the cooling devices contributed towards bird welfare by altering the behavioral expression from agonistic to non-agonistic activities.RNA-guided recombinases (RGR) are potentially valuable tools for basic research and genetic modifications. The platform has been demonstrated to do genome editing efficiently. The platform operates on a typical recognition site comprised of the degenerate recombinase site, a 5 to 6-base pair spacer flanking it and this whole central region is flanked by two guide RNA-specified DNA sequences or Cas9 binding sites which is followed by protospacer adjacent motifs. In present investigation, a detailed map of target sites for RNA-guided recombinase platforms based on hyperactivated recombinase Beta throughout the bovine genome was prepared. For this, Chromosome wise whole genomic sequence data was retrieved from Ensembl followed by designing search pattern for recombinase Beta with spacer length five. By using this search pattern, RGR target sites were located by using dreg program of Emboss package. In total,436 RGR target sites were identified in bovine genome for recombinase Beta with spacer length five. These RGR target site provide potential of being utilized for specific genomic integration, deletion or inversion.