CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>

ABSTRACT Sand flies are the natural vectors for the Leishmania species that produce a spectrum of diseases in their mammalian hosts, including humans. Studies of sand fly/Leishmania interactions have been limited by the absence of genome editing techniques applicable to these insects. In this report...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Isabelle Louradour, Kashinath Ghosh, Ehud Inbar, David L. Sacks
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	CRISPR/Cas9 IMD pathway Leishmania sand fly Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/33eb51800b104e918f96c02481bb267c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:33eb51800b104e918f96c02481bb267c
record_format	dspace
spelling	oai:doaj.org-article:33eb51800b104e918f96c02481bb267c2021-11-15T16:22:09ZCRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>10.1128/mBio.01941-192150-7511https://doaj.org/article/33eb51800b104e918f96c02481bb267c2019-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01941-19https://doaj.org/toc/2150-7511ABSTRACT Sand flies are the natural vectors for the Leishmania species that produce a spectrum of diseases in their mammalian hosts, including humans. Studies of sand fly/Leishmania interactions have been limited by the absence of genome editing techniques applicable to these insects. In this report, we adapted CRISPR (clustered regularly interspaced palindromic repeat)/Cas9 (CRISPR-associated protein 9) technology to the Phlebotomus papatasi sand fly, a natural vector for Leishmania major, targeting the sand fly immune deficiency (IMD) pathway in order to decipher its contribution to vector competence. We established a protocol for transformation in P. papatasi and were able to generate transmissible null mutant alleles for Relish (Rel), the only transcription factor of the IMD pathway. While the maintenance of a homozygous mutant stock was severely compromised, we were able to establish in an early generation their greater susceptibility to infection with L. major. Flies carrying different heterozygous mutant alleles variably displayed a more permissive phenotype, presenting higher loads of parasites or greater numbers of infective-stage promastigotes. Together, our data show (i) the successful adaptation of the CRISPR/Cas9 technology to sand flies and (ii) the impact of the sand fly immune response on vector competence for Leishmania parasites. IMPORTANCE Sand flies are the natural vectors of Leishmania parasites. Studies of sand fly/Leishmania interactions have been limited by the lack of successful genomic manipulation of these insects. This paper shows the first example of successful targeted mutagenesis in sand flies via adaptation of the CRISPR/Cas9 editing technique. We generated transmissible null mutant alleles of relish, a gene known to be essential for the control of immune response in other insects. In addition to the expected higher level of susceptibility to bacteria, the mutant flies presented higher loads of parasites when infected with L. major, showing that the sand fly immune response impacts its vector competence for this pathogen.Isabelle LouradourKashinath GhoshEhud InbarDavid L. SacksAmerican Society for MicrobiologyarticleCRISPR/Cas9IMD pathwayLeishmaniasand flyMicrobiologyQR1-502ENmBio, Vol 10, Iss 4 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPR/Cas9 IMD pathway Leishmania sand fly Microbiology QR1-502
spellingShingle	CRISPR/Cas9 IMD pathway Leishmania sand fly Microbiology QR1-502 Isabelle Louradour Kashinath Ghosh Ehud Inbar David L. Sacks CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>
description	ABSTRACT Sand flies are the natural vectors for the Leishmania species that produce a spectrum of diseases in their mammalian hosts, including humans. Studies of sand fly/Leishmania interactions have been limited by the absence of genome editing techniques applicable to these insects. In this report, we adapted CRISPR (clustered regularly interspaced palindromic repeat)/Cas9 (CRISPR-associated protein 9) technology to the Phlebotomus papatasi sand fly, a natural vector for Leishmania major, targeting the sand fly immune deficiency (IMD) pathway in order to decipher its contribution to vector competence. We established a protocol for transformation in P. papatasi and were able to generate transmissible null mutant alleles for Relish (Rel), the only transcription factor of the IMD pathway. While the maintenance of a homozygous mutant stock was severely compromised, we were able to establish in an early generation their greater susceptibility to infection with L. major. Flies carrying different heterozygous mutant alleles variably displayed a more permissive phenotype, presenting higher loads of parasites or greater numbers of infective-stage promastigotes. Together, our data show (i) the successful adaptation of the CRISPR/Cas9 technology to sand flies and (ii) the impact of the sand fly immune response on vector competence for Leishmania parasites. IMPORTANCE Sand flies are the natural vectors of Leishmania parasites. Studies of sand fly/Leishmania interactions have been limited by the lack of successful genomic manipulation of these insects. This paper shows the first example of successful targeted mutagenesis in sand flies via adaptation of the CRISPR/Cas9 editing technique. We generated transmissible null mutant alleles of relish, a gene known to be essential for the control of immune response in other insects. In addition to the expected higher level of susceptibility to bacteria, the mutant flies presented higher loads of parasites when infected with L. major, showing that the sand fly immune response impacts its vector competence for this pathogen.
format	article
author	Isabelle Louradour Kashinath Ghosh Ehud Inbar David L. Sacks
author_facet	Isabelle Louradour Kashinath Ghosh Ehud Inbar David L. Sacks
author_sort	Isabelle Louradour
title	CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>
title_short	CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>
title_full	CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>
title_fullStr	CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>
title_full_unstemmed	CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>
title_sort	crispr/cas9 mutagenesis in <named-content content-type="genus-species">phlebotomus papatasi</named-content>: the immune deficiency pathway impacts vector competence for <named-content content-type="genus-species">leishmania major</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/33eb51800b104e918f96c02481bb267c
work_keys_str_mv	AT isabellelouradour crisprcas9mutagenesisinnamedcontentcontenttypegenusspeciesphlebotomuspapatasinamedcontenttheimmunedeficiencypathwayimpactsvectorcompetencefornamedcontentcontenttypegenusspeciesleishmaniamajornamedcontent AT kashinathghosh crisprcas9mutagenesisinnamedcontentcontenttypegenusspeciesphlebotomuspapatasinamedcontenttheimmunedeficiencypathwayimpactsvectorcompetencefornamedcontentcontenttypegenusspeciesleishmaniamajornamedcontent AT ehudinbar crisprcas9mutagenesisinnamedcontentcontenttypegenusspeciesphlebotomuspapatasinamedcontenttheimmunedeficiencypathwayimpactsvectorcompetencefornamedcontentcontenttypegenusspeciesleishmaniamajornamedcontent AT davidlsacks crisprcas9mutagenesisinnamedcontentcontenttypegenusspeciesphlebotomuspapatasinamedcontenttheimmunedeficiencypathwayimpactsvectorcompetencefornamedcontentcontenttypegenusspeciesleishmaniamajornamedcontent
_version_	1718426895655108608

CRISPR/Cas9 Mutagenesis in <named-content content-type="genus-species">Phlebotomus papatasi</named-content>: the Immune Deficiency Pathway Impacts Vector Competence for <named-content content-type="genus-species">Leishmania major</named-content>

Ejemplares similares