Cargando…

CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken

Salmonella Gallinarum causes fowl typhoid in poultry leading to a huge economic loss to the poultry industry. The large virulence plasmid of S. gallinarum has been associated with various systemic infections in poultry. A five-gene spanning region (spvRABCD) of 7.8 kb on the large plasmid mainly con...

Descripción completa

Detalles Bibliográficos
Autores principales: Basit, Abdul, Tahir, Hamza, Haider, Zulquernain, Tariq, Hafsa, Ullah, Asim, Rehman, Shafiq Ur
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9234527/
https://www.ncbi.nlm.nih.gov/pubmed/35769104
http://dx.doi.org/10.3389/fbioe.2022.885227
_version_ 1784736096626671616
author Basit, Abdul
Tahir, Hamza
Haider, Zulquernain
Tariq, Hafsa
Ullah, Asim
Rehman, Shafiq Ur
author_facet Basit, Abdul
Tahir, Hamza
Haider, Zulquernain
Tariq, Hafsa
Ullah, Asim
Rehman, Shafiq Ur
author_sort Basit, Abdul
collection PubMed
description Salmonella Gallinarum causes fowl typhoid in poultry leading to a huge economic loss to the poultry industry. The large virulence plasmid of S. gallinarum has been associated with various systemic infections in poultry. A five-gene spanning region (spvRABCD) of 7.8 kb on the large plasmid mainly confers virulence to the bacteria. However, the exact role of these genes in virulence has not been elucidated yet. SpvB exhibits delayed cell death by preventing actin polymerization followed by apoptosis during intracellular infection. The specific role of SpvB in causing the disease is not known yet. In the current study, the SpvB gene was deleted through CRISPR/Cas9 method from a large virulent plasmid of locally isolated S. gallinarum strain (SG18). The homology-directed repair method was used for complete deletion of SpvB gene using the modified pCas9 plasmid. The SpvB-deleted S. gallinarum strain (ΔSpvB_SG18), when tested for its virulence in broiler chicken showed no diseases signs and mortality. In addition, the avirulent strain does not affect the bird’s weight and was rapidly cleared from the liver after infection. However, it cleared from the intestine only after 4–5 days, which suggests that the ΔSpvB_SG18 strain is unable to invade from the intestine to the liver. This is the first study to report a complete gene deletion from the S. gallinarum virulent plasmid and its effect. This method will be useful for the deletion of virulent genes from S. gallinarum, to study their role in pathogenesis, and to prepare an effective vaccine strain for controlling fowl typhoid in poultry.
format Online
Article
Text
id pubmed-9234527
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-92345272022-06-28 CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken Basit, Abdul Tahir, Hamza Haider, Zulquernain Tariq, Hafsa Ullah, Asim Rehman, Shafiq Ur Front Bioeng Biotechnol Bioengineering and Biotechnology Salmonella Gallinarum causes fowl typhoid in poultry leading to a huge economic loss to the poultry industry. The large virulence plasmid of S. gallinarum has been associated with various systemic infections in poultry. A five-gene spanning region (spvRABCD) of 7.8 kb on the large plasmid mainly confers virulence to the bacteria. However, the exact role of these genes in virulence has not been elucidated yet. SpvB exhibits delayed cell death by preventing actin polymerization followed by apoptosis during intracellular infection. The specific role of SpvB in causing the disease is not known yet. In the current study, the SpvB gene was deleted through CRISPR/Cas9 method from a large virulent plasmid of locally isolated S. gallinarum strain (SG18). The homology-directed repair method was used for complete deletion of SpvB gene using the modified pCas9 plasmid. The SpvB-deleted S. gallinarum strain (ΔSpvB_SG18), when tested for its virulence in broiler chicken showed no diseases signs and mortality. In addition, the avirulent strain does not affect the bird’s weight and was rapidly cleared from the liver after infection. However, it cleared from the intestine only after 4–5 days, which suggests that the ΔSpvB_SG18 strain is unable to invade from the intestine to the liver. This is the first study to report a complete gene deletion from the S. gallinarum virulent plasmid and its effect. This method will be useful for the deletion of virulent genes from S. gallinarum, to study their role in pathogenesis, and to prepare an effective vaccine strain for controlling fowl typhoid in poultry. Frontiers Media S.A. 2022-06-13 /pmc/articles/PMC9234527/ /pubmed/35769104 http://dx.doi.org/10.3389/fbioe.2022.885227 Text en Copyright © 2022 Basit, Tahir, Haider, Tariq, Ullah and Rehman. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Basit, Abdul
Tahir, Hamza
Haider, Zulquernain
Tariq, Hafsa
Ullah, Asim
Rehman, Shafiq Ur
CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken
title CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken
title_full CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken
title_fullStr CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken
title_full_unstemmed CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken
title_short CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken
title_sort crispr/cas9-based deletion of spvb gene from salmonella gallinarum leads to loss of virulence in chicken
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9234527/
https://www.ncbi.nlm.nih.gov/pubmed/35769104
http://dx.doi.org/10.3389/fbioe.2022.885227
work_keys_str_mv AT basitabdul crisprcas9baseddeletionofspvbgenefromsalmonellagallinarumleadstolossofvirulenceinchicken
AT tahirhamza crisprcas9baseddeletionofspvbgenefromsalmonellagallinarumleadstolossofvirulenceinchicken
AT haiderzulquernain crisprcas9baseddeletionofspvbgenefromsalmonellagallinarumleadstolossofvirulenceinchicken
AT tariqhafsa crisprcas9baseddeletionofspvbgenefromsalmonellagallinarumleadstolossofvirulenceinchicken
AT ullahasim crisprcas9baseddeletionofspvbgenefromsalmonellagallinarumleadstolossofvirulenceinchicken
AT rehmanshafiqur crisprcas9baseddeletionofspvbgenefromsalmonellagallinarumleadstolossofvirulenceinchicken