دورية أكاديمية

Coupling CRISPR/Cas9 and Lambda Red Recombineering System for Genome Editing of Salmonella Gallinarum and the Effect of ssaU Knock-Out Mutant on the Virulence of Bacteria

التفاصيل البيبلوغرافية
العنوان: Coupling CRISPR/Cas9 and Lambda Red Recombineering System for Genome Editing of Salmonella Gallinarum and the Effect of ssaU Knock-Out Mutant on the Virulence of Bacteria
المؤلفون: Hamza Tahir, Abdul Basit, Hafsa Tariq, Zulquernain Haider, Asim Ullah, Zafar Hayat, Shafiq Ur Rehman
المصدر: Biomedicines, Vol 10, Iss 12, p 3028 (2022)
بيانات النشر: MDPI AG, 2022.
سنة النشر: 2022
المجموعة: LCC:Biology (General)
مصطلحات موضوعية: CRISPR/Cas9, lambda recombineering, genome editing, type three secretion system, virulent genes, Salmonella Gallinarum, Biology (General), QH301-705.5
الوصف: The poultry industry in developing countries still faces a significant threat from fowl typhoid, a disease caused by Salmonella Gallinarum that has been well contained in more economically developed countries. In addition to the virulence exhibited by large virulence plasmid (85 kb), Salmonella Pathogenicity Island 2 in S. Gallinarum plays a key role in mediating disease through its type III secretion systems (TTSS). The TTSS secrete effector protein across the Salmonella containing vacuoles and mediate the internalization of bacteria by modulating vesicular passage. In this study, candidate virulent ssaU gene (~1 kb) encoding type III secretion system was successfully deleted from indigenously isolated S. Gallinarum genome through homology-directed repair using CRISPR/Cas9 and lambda recombination systems. CRISPR/Cas9-based genome editing of poultry-derived Salmonella Gallinarum has not been previously reported, which might be linked to a lack of efficiency in its genetic tools. This is the first study which demonstrates a complete CRISPR/Cas9-based gene deletion from this bacterial genome. More importantly, a poultry experimental model was employed to assess the virulence potential of this mutant strain (ΔssaU_SG18) which was unable to produce any mortality in the experimentally challenged birds as compared to the wild type strain. No effect on weight gain was observed whereas bacteria were unable to colonize the intestine and liver in our challenge model. This in vivo loss of virulence in mutant strain provides an excellent functionality of this system to be useful in live vaccine development against this resistant and patho genic bacteria.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2227-9059
Relation: https://www.mdpi.com/2227-9059/10/12/3028; https://doaj.org/toc/2227-9059
DOI: 10.3390/biomedicines10123028
URL الوصول: https://doaj.org/article/c89d0f1dda184e93b014290fc808b0b3
رقم الأكسشن: edsdoj.89d0f1dda184e93b014290fc808b0b3
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:22279059
DOI:10.3390/biomedicines10123028