S Anandu¹, Basavaraj Sajjanar², Vijayakumar Jawalagatti³, Laharika Reddy¹, Yashica KA¹, Saravanan BC⁴ and Anup Kumar Tewari¹*

¹Division of Parasitology, ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
²Division of Animal Biotechnology, ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
³Mayo Clinic, Rochester, United States
⁴CADRAD, ICAR- ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India

*Corresponding Author: Anup Kumar Tewari, Division of Parasitology, ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.

Received: November 26, 2024; ; Published: January 31, 2025

Abstract

Trypanosoma evansi, a blood protozoan parasite of high economic importance, has a wide prevalence in the vertebrates in many tropical countries including India. The parasite is transmitted by the hematophagous flies and causes a chronic debilitating disease ‘Surra’ in domestic livestock. Development of a protective vaccine has not yet been possible due to the ability of the parasite to evade the host's immune response by periodically changing its surface glycoproteins. The Paraflagellar Rod 1 (PFR1) gene codes for a major flagellar constituent PFR1 protein of T. evansi. We report here a specifically targeted editing of the T. evansi PFR1 gene by the CRISPR Cas9 tool. We combined the single guide RNA (sgRNA) and recombinant Cas9 protein in-vitro to form a ribonucleoprotein complex (RNP) for transfecting T. evansi and the induced mutation in the coding sequence of PFR1 was assessed by T7E1 endonuclease assay. The CRISPR-Cas9-mediated PFR1 gene-edited parasites had a significantly shorter lifespan in vitro.

 Keywords: Trypanosoma Evansi; PFR1; CRISPR; Cas9; Gene Editing

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Citation

Citation: Anup Kumar Tewari., et al. “CRISPR-Cas9 Mediated Paraflagellar rod 1 Gene Editing in Trypanosoma Evansi". Acta Scientific Veterinary Sciences 7.2 (2025): 33-39.

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Copyright: © 2025 Anup Kumar Tewari., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.