Study of mRNA expression levels in steady state Trypanosoma evansi for plausible selection of housekeeping and drug target genes Gupta Snehil, Vohra Sukhdeep, Kumar Rajender1,* Department of Veterinary Parasitology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125004, India 1Parasitology Lab, ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India *Corresponding author. Email: rkg.nrce@gmail.com
Online Published on 15 February, 2024. Abstract Surra is a dreadful infectious protozoan disease affecting many animal hosts. Emerging drug resistance, a limited number of drugs, and their high cytotoxicity have motivated researchers to select novel and safe target-specific therapeutics against Trypanosoma evansi. In the absence of transcriptomic data and limited genomic information, selecting housekeeping genes and drug targets is challenging. Therefore, the present study aimed for comparative analysis of mRNA expression levels of various genes in steady state T. evansi for plausible selection of housekeeping and drug target genes. The fully adapted and stable T. evansi (equine isolate) population under in vitro conditions (HMI-9 media) was used for RNA isolation. Sequence similarity between T. evansi and related trypanosomatid species was used as a computational approach to design primers for several genes, unexplored earlier in this organism. We have quantified the mRNA expression level of seven housekeeping genes and 13 drug target genes in T. evansi using one-step qPCR. Out of the seven candidate housekeeping genes, β-tubulin was having highest copy number (Ct, 15.00 ± 0.15), whereas, telomerase reverse transcriptase (Ct, 19.55 ± 0.17) and double-strand break repair nuclease (Ct, 19.55 ± 0.13) genes had lowest copy number. Out of the 13 drug target genes, arginine kinase was found to be the most promising with the highest copy number (Ct, 17.92 ± 0.09), however, hexokinase had least (Ct, 28.20 ± 0.26) mRNA expression levels. Data generated showed that qPCR can be employed to select drug target genes. The information can further be utilized for the development of novel drug molecules as well as vaccine candidates for curtailment of Surra in livestock animals. Top Keywords Trypanosoma evansi, qPCR, Housekeeping genes, Drug targets, HMI-9 medium. Top |