Journal of Vector Borne Diseases

RESEARCH ARTICLE
Year
: 2019  |  Volume : 56  |  Issue : 2  |  Page : 98--104

Transient knockdown of Nucleoside transporter 4 gene expression as a therapeutic target in Leishmania major by antisense RNA: In vitro and in vivo studies


Farideh Tohidi1, Zahra Babaei2, Bahram Kazemi3, Mojgan Bandehpour3, Iraj Sharifi2, Mohammad Reza Rabiei4, Ebrahim Saedi Dezaki5 
1 Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman; Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran
2 Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran
3 Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
4 Department of Statistics, Faculty of Mathematical Sciences, Shahrood University of Technology, Shahrood, Iran
5 Department of Medical Parasitology and Mycology, School of Medicine, Shahrekord University of Medical Sciences, Shahrekord, Iran

Correspondence Address:
Zahra Babaei
Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman
Iran

Background & objectives: Leishmania parasites cause various clinical symptoms in humans such as cutaneous ulcers and fatal visceral diseases. These parasites cannot synthesize purine rings de novo and must uptake purines from their hosts via salvage. Salvage is regulated by permeases in the cell membrane. There are hundreds of membrane transporter proteins to receive nutrients in Leishmania. Nucleoside transporter 4 (NT4) is one of the purine transporters that is involved in enhancing the uptake of adenine in Leishmania major. They are important new drug targets for the treatment of leishmaniasis because they can be used to transport toxic purine analogs to kill parasitic cells, thus preventing the progression of the infection. The present study was conducted to silence the NT4 nucleobase involved in the salvage pathway to interrupt purine nucleotide membrane transport in the cells of L. major. Methods: In this study, a 502 bp segment of NT4 gene sequence was selected and designed as antisense transcripts after insertion in the parasite. The NT4 construct was transfected into L. major promastigotes for in vitro study of gene expression. Then, BALB/c mice infected with transgenic Leishmania and wild-type strain along with the number and size of lesions were studied in vivo. Results: The study showed that relative expression of NT4 gene in mutant Leishmania was lower than in the control on Day 3 to 20. The percentages and the number of amastigotes in infected macrophages with wild-type strain L. major were more than infected macrophages with mutant parasites. Infected BALB/c mice with transgenic Leish- mania showed a lower number and size of lesions than the BALB/c mice infected with wild-type strain. Interpretation & conclusion: The results of the study indicated that the use of antisense RNA reduced NT4 gene expression in L. major. Further, studies are needed to ascertain that the use of antisense can be considered as a new treatment for leishmaniasis.


How to cite this article:
Tohidi F, Babaei Z, Kazemi B, Bandehpour M, Sharifi I, Rabiei MR, Dezaki ES. Transient knockdown of Nucleoside transporter 4 gene expression as a therapeutic target in Leishmania major by antisense RNA: In vitro and in vivo studies.J Vector Borne Dis 2019;56:98-104


How to cite this URL:
Tohidi F, Babaei Z, Kazemi B, Bandehpour M, Sharifi I, Rabiei MR, Dezaki ES. Transient knockdown of Nucleoside transporter 4 gene expression as a therapeutic target in Leishmania major by antisense RNA: In vitro and in vivo studies. J Vector Borne Dis [serial online] 2019 [cited 2019 Sep 19 ];56:98-104
Available from: http://www.jvbd.org/article.asp?issn=0972-9062;year=2019;volume=56;issue=2;spage=98;epage=104;aulast=Tohidi;type=0