The genetic toolbox for<i>Leishmania</i>parasites

Roberts, Sigrid C.

doi:10.4161/bbug.2.6.18205

Cited by 12 publications

(14 citation statements)

References 70 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Advances in genetic manipulation of Leishmania parasites has considerably facilitated the characterisation of metabolic processes and molecules important for parasite proliferation or virulence [ 3 , 4 ]. Nevertheless, the study of essential genes remains difficult since knockouts can only be performed if rescue strategies such as nutrient supplementation or ectopic gene copies can be used.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Depletion of UDP-Glucose and UDP-Galactose Using a Degron System Leads to Growth Cessation of Leishmania major

et al. 2015

View full text Add to dashboard Cite

Interconversion of UDP-glucose (UDP-Glc) and UDP-galactose (UDP-Gal) by the UDP-Glc 4´-epimerase intimately connects the biosynthesis of these two nucleotide sugars. Their de novo biosynthesis involves transformation of glucose-6-phosphate into glucose-1-phosphate by the phosphoglucomutase and subsequent activation into UDP-Glc by the specific UDP-Glc pyrophosphorylase (UGP). Besides UGP, Leishmania parasites express an uncommon UDP-sugar pyrophosphorylase (USP) able to activate both galactose-1-phosphate and glucose-1-phosphate in vitro. Targeted gene deletion of UGP alone was previously shown to principally affect expression of lipophosphoglycan, resulting in a reduced virulence. Since our attempts to delete both UGP and USP failed, deletion of UGP was combined with conditional destabilisation of USP to control the biosynthesis of UDP-Glc and UDP-Gal. Stabilisation of the enzyme produced by a single USP allele was sufficient to maintain the steady-state pools of these two nucleotide sugars and preserve almost normal glycoinositolphospholipids galactosylation, but at the apparent expense of lipophosphoglycan biosynthesis. However, under destabilising conditions, the absence of both UGP and USP resulted in depletion of UDP-Glc and UDP-Gal and led to growth cessation and cell death, suggesting that either or both of these metabolites is/are essential.

show abstract

Section: Introductionmentioning

confidence: 99%

“…major and L . donovani lack a functional RNAi pathway [ 4 ]. More recently, a system for conditional destabilisation of protein has been described.…”

Section: Introductionmentioning

confidence: 99%

Depletion of UDP-Glucose and UDP-Galactose Using a Degron System Leads to Growth Cessation of Leishmania major

et al. 2015

View full text Add to dashboard Cite

show abstract

“…1 ). When an episomal vector was used, it is possible that the daughter cells do not receive equal amounts of the vector, and the expression level of genes may vary from cell to cell, allowing the mosaicism observed 40 .…”

Section: Discussionmentioning

confidence: 99%

Growth arrested live-attenuated Leishmania infantum KHARON1 null mutants display cytokinesis defect and protective immunity in mice

Santi

Lanza

Tunes

et al. 2018

Sci Rep

View full text Add to dashboard Cite

There is no safe and efficacious vaccine against human leishmaniasis available and live attenuated vaccines have been used as a prophylactic alternative against the disease. In order to obtain an attenuated Leishmania parasite for vaccine purposes, we generated L. infantum KHARON1 (KH1) null mutants (ΔLikh1). This gene was previously associated with growth defects in L. mexicana. ΔLikh1 was obtained and confirmed by PCR, qPCR and Southern blot. We also generate a KH1 complemented line with the introduction of episomal copies of KH1. Although ΔLikh1 promastigote forms exhibited a growth pattern similar to the wild-type line, they differ in morphology without affecting parasite viability. L. infantum KH1-deficient amastigotes were unable to sustain experimental infection in macrophages, forming multinucleate cells which was confirmed by in vivo attenuation phenotype. The cell cycle analysis of ΔLikh1 amastigotes showed arrested cells at G2/M phase. ΔLikh1-immunized mice presented reduced parasite burden upon challenging with virulent L. infantum, when compared to naïve mice. An effect associated with increased Li SLA-specific IgG serum levels and IL-17 production. Thus, ΔLikh1 parasites present an infective-attenuated phenotype due to a cytokinesis defect, whereas it induces immunity against visceral leishmaniasis in mouse model, being a candidate for antileishmanial vaccine purposes.

show abstract

“…Na busca por explicações sobre mecanismos e fatores de virulência e sua relação com o desenvolvimento das leishmanioses, nos últimos anos uma variedade de técnicas de manipulação genética (Beverley, 2003a;Roberts, 2011), em associação com os resultados dos projetos de genoma de várias espécies de Leishmania (Downing et al, 2011;Ivens et al, 2005;Llanes et al, 2015;Peacock et al, 2007;Rogers et al, 2011), permitiram ampliar o conhecimento sobre a biologia desses parasitos e sua interação parasito-hospedeiro. Análises comparativas entre os genomas revelaram um alto grau de conservação no conteúdo de genes e arquitetura entre as espécies do subgênero Viannia e…”

Section: Estudos Sobre Virulência De Leishmania Por Genética Reversaunclassified

“…Os genes associados a virulência têm um profundo impacto na biologia do parasita, por isso são considerados alvos potenciais de vacinação ou quimioterapia (Duncan et al, 2011 (Beverley, 2003b;Roberts, 2011). Todas estas técnicas representam formas robustas de caracterização genética que permitem identificar e investigar genes associados a virulência (Alexandratos et al, 2013;Antoniazi et al, 2000;Zhang and Matlashewski, 2010) ou estudar a regulação da expressão gênica (Brooks et al, 2001;Lahav et al, 2011;Mishra et al, 2003); identificar genes essenciais para a sobrevivência Introdução (Cruz et al, 1991;Cruz et al, 1993;Hassan et al, 2001;Price et al, 2003) ou envolvidos nos mecanismos de aquisição de resistência a drogas (Coelho et al, 2003;Ouellette et al, 1998).…”

Section: Estudos Sobre Virulência De Leishmania Por Genética Reversaunclassified

Investigação de genes potencialmente implicados no controle de virulência em <i>Leishmania major</i>

Londoño¹

View full text Add to dashboard Cite

show abstract

The genetic toolbox forLeishmaniaparasites

Cited by 12 publications

References 70 publications

Depletion of UDP-Glucose and UDP-Galactose Using a Degron System Leads to Growth Cessation of Leishmania major

Depletion of UDP-Glucose and UDP-Galactose Using a Degron System Leads to Growth Cessation of Leishmania major

Growth arrested live-attenuated Leishmania infantum KHARON1 null mutants display cytokinesis defect and protective immunity in mice

Investigação de genes potencialmente implicados no controle de virulência em <i>Leishmania major</i>

Contact Info

Product

Resources

About