title: The role of GlcNAc-PI-de-N-acetylase gene by gene knockout through homologous recombination and its consequences on survival, growth and infectivity of Leishmania major in in vitro and in vivo conditions.
abstract: At present, there are no efficacious vaccines or effective drugs against leishmaniasis; therefore new and innovative control methods are urgently required. One way to achieve this important goal is through using reverse genetic engineering to evaluate important enzymes, proteins and macromolecules. One of the most important enzymes for Glycosylphosphatidylinositol (GPI) biosynthetic pathways is GlcNAc-PI-de-N-acetylase (GPI12). The molecular constructs were cloned in Escherichia coli strain Top 10 and confirmed by molecular methods and were transfected by electroporation into Leishmania major. We demonstrated that two alleles of the GPI12 gene in L. major were successfully removed and enabling the generation of a null mutant, which supports the idea that GPI12 is not an essential gene for the growth and survival of Leishmania and the homozygous knockouts of Leishmania are able to survive. We were able to produce a mutant parasite that caused no damaged to the host. Further investigations are essential to check the safety profile in laboratory animals.
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Pooya Ghasemi Nejad
authors.initials: PG
authors.lastname: Almani
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; Leishmaniasis Research Centre, Kerman University of Medical Sciences, Kerman, Iran. Electronic address: iraj.sharifi@yahoo.com.
authors.forename: Iraj
authors.initials: I
authors.lastname: Sharifi
authors.affiliation: Cellular and Molecular Biology, Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Biotechnology Department, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Iran. Electronic address: bahram_14@yahoo.com.
authors.forename: Bahram
authors.initials: B
authors.lastname: Kazemi
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; Leishmaniasis Research Centre, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Babaei
authors.affiliation: Cellular and Molecular Biology, Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Biotechnology Department, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Iran.
authors.forename: Mojgan
authors.initials: M
authors.lastname: Bandehpour
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Samira
authors.initials: S
authors.lastname: Salari
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Ebrahim Saedi
authors.initials: ES
authors.lastname: Dezaki
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; Department of Medical Parasitology and Mycology, School of Medicine, Golestan University of Medical Sciences, Golestan, Iran.
authors.forename: Farideh
authors.initials: F
authors.lastname: Tohidi
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Mohammadi
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D005260:Female; D055786:Gene Knockout Techniques; D005838:Genotype; D059765:Homologous Recombination; D018320:Leishmania major / Q000472:pathogenicity / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000581:Amidohydrolases; C087696:N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase
keywords: GPI12; Leishmaniasis; Null mutant; Reverse genetic engineering