Development of a CRISPR/Cas9 system against ruminant animal brucellosis
Abstract Background Brucellosis, caused by several Brucella species, such as the bacterium Brucella melitensis, is considered one of the most severe zoonotic diseases worldwide. Not only does it affect ruminant animal populations, leading to a substantial financial burden for stockbreeders, but also...
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | BMC Veterinary Research |
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| Online Access: | http://link.springer.com/article/10.1186/s12917-019-2179-z |
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| author | Garyfalia Karponi Spyridon K. Kritas Gina Papadopoulou Elissavet-Kalliopi Akrioti Eleni Papanikolaou Evanthia Petridou |
| author_facet | Garyfalia Karponi Spyridon K. Kritas Gina Papadopoulou Elissavet-Kalliopi Akrioti Eleni Papanikolaou Evanthia Petridou |
| author_sort | Garyfalia Karponi |
| collection | DOAJ |
| description | Abstract Background Brucellosis, caused by several Brucella species, such as the bacterium Brucella melitensis, is considered one of the most severe zoonotic diseases worldwide. Not only does it affect ruminant animal populations, leading to a substantial financial burden for stockbreeders, but also poses severe public health issues. For almost four decades in southern Europe and elsewhere, eradication of the disease has been based on ambiguously effective programs, rendering massive sanitation of livestock urgent and indispensable. Gene therapy, which has been proved effective in the clinic, could possibly constitute an alternative option towards a permanent cure for brucellosis, by aiding in the deletion or inactivation of genes associated with the replication of Brucella within the host cells. Results We infected ovine macrophages with B.melitensis, to simulate the host cell/microorganism interaction in vitro, and transduced the infected cells with CRISPR/Cas9 lentiviral vectors that target Brucella’s RNA polymerase subunit A (RpolA) or virulence-associated gene virB10 at a multiplicity of infection of 60. We demonstrate a significant decrease in the bacterial load per cell when infected cells are transduced with the RpolA vector and that the number of internalized brucellae per cell remains unaffected when macrophages are transduced with a conventional lentiviral vector expressing the green fluorescence protein, thus underlining the bactericidal effect of our CRISPR/Cas9 system. Conclusions Pending in vivo verification of our findings, overall, these results may prove critical not only for the treatment of human brucellosis, but for other infectious diseases in general. |
| first_indexed | 2024-12-19T09:06:17Z |
| format | Article |
| id | doaj.art-8c978276bf5f4da5b832bd8f2c6b59dd |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-19T09:06:17Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-8c978276bf5f4da5b832bd8f2c6b59dd2022-12-21T20:28:20ZengBMCBMC Veterinary Research1746-61482019-11-0115111010.1186/s12917-019-2179-zDevelopment of a CRISPR/Cas9 system against ruminant animal brucellosisGaryfalia Karponi0Spyridon K. Kritas1Gina Papadopoulou2Elissavet-Kalliopi Akrioti3Eleni Papanikolaou4Evanthia Petridou5Department of Microbiology and Infectious Diseases, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of ThessalonikiDepartment of Microbiology and Infectious Diseases, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of ThessalonikiCellular Immunology Laboratory, Basic Research, Biomedical Research Foundation of the Academy of AthensLaboratory of Cellular and Molecular Neurobiology-Stem Cells, Hellenic Pasteur InstituteLaboratory of Biology, School of Medicine, National and Kapodistrian University of AthensDepartment of Microbiology and Infectious Diseases, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of ThessalonikiAbstract Background Brucellosis, caused by several Brucella species, such as the bacterium Brucella melitensis, is considered one of the most severe zoonotic diseases worldwide. Not only does it affect ruminant animal populations, leading to a substantial financial burden for stockbreeders, but also poses severe public health issues. For almost four decades in southern Europe and elsewhere, eradication of the disease has been based on ambiguously effective programs, rendering massive sanitation of livestock urgent and indispensable. Gene therapy, which has been proved effective in the clinic, could possibly constitute an alternative option towards a permanent cure for brucellosis, by aiding in the deletion or inactivation of genes associated with the replication of Brucella within the host cells. Results We infected ovine macrophages with B.melitensis, to simulate the host cell/microorganism interaction in vitro, and transduced the infected cells with CRISPR/Cas9 lentiviral vectors that target Brucella’s RNA polymerase subunit A (RpolA) or virulence-associated gene virB10 at a multiplicity of infection of 60. We demonstrate a significant decrease in the bacterial load per cell when infected cells are transduced with the RpolA vector and that the number of internalized brucellae per cell remains unaffected when macrophages are transduced with a conventional lentiviral vector expressing the green fluorescence protein, thus underlining the bactericidal effect of our CRISPR/Cas9 system. Conclusions Pending in vivo verification of our findings, overall, these results may prove critical not only for the treatment of human brucellosis, but for other infectious diseases in general.http://link.springer.com/article/10.1186/s12917-019-2179-zBrucellosisGene therapyViral vectorsRuminant animalsMacrophages |
| spellingShingle | Garyfalia Karponi Spyridon K. Kritas Gina Papadopoulou Elissavet-Kalliopi Akrioti Eleni Papanikolaou Evanthia Petridou Development of a CRISPR/Cas9 system against ruminant animal brucellosis BMC Veterinary Research Brucellosis Gene therapy Viral vectors Ruminant animals Macrophages |
| title | Development of a CRISPR/Cas9 system against ruminant animal brucellosis |
| title_full | Development of a CRISPR/Cas9 system against ruminant animal brucellosis |
| title_fullStr | Development of a CRISPR/Cas9 system against ruminant animal brucellosis |
| title_full_unstemmed | Development of a CRISPR/Cas9 system against ruminant animal brucellosis |
| title_short | Development of a CRISPR/Cas9 system against ruminant animal brucellosis |
| title_sort | development of a crispr cas9 system against ruminant animal brucellosis |
| topic | Brucellosis Gene therapy Viral vectors Ruminant animals Macrophages |
| url | http://link.springer.com/article/10.1186/s12917-019-2179-z |
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