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Reprogramming Mycobacterium tuberculosis CRISPR System for Gene Editing and Genomewide RNA Interference Screening

Genomics, Proteomics & Bioinformatics 2022年第6期20卷 1180-1196页

作者： Khaista Rahman muhammad Jamal Xi Chen Wei Zhou Bin Yang Yanyan Zou Weize Xu Yingying Lei Chengchao Wu Xiaojian Cao Rohit Tyagi muhammad ahsan naeem Da Lin Zeshan Habib Nan Peng Zhen F.Fu Gang Cao State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhan 430070China College of Veterinary Medicine Huazhong Agricultural UniversityWuhan 430070China Bio-Medical Center Huazhong Agricultural UniversityWuhan 430070China Key Laboratory of Preventive Veterinary Medicine in Hubei Province the Cooperative Innovation Center for Sustainable Pig ProductionWuhan 430070China

Mycobacterium tuberculosis is the causative agent of tuberculosis(TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M. tuberculosis. Here, we harnessed an endogenous type Ⅲ-A CRISPR/Cas10 system of M. tuberculosis for efficient gene editing and RNA interference(RNAi).This simple and easy method only needs to transform a single mini-CRISPR array plasmid, thus avoiding the introduction of exogenous protein and minimizing proteotoxicity. We demonstrated that M. tuberculosis genes can be efficiently and specifically knocked in/out by this system as confirmed by DNA high-throughput sequencing. This system was further applied to single-and multiple-gene RNAi. Moreover, we successfully performed genome-wide RNAi screening to identify M. tuberculosis genes regulating in vitro and intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.

关键词： Mycobacterium tuberculosis Type III-A CRISPR system Gene editing Gene interference Genome-wide RNAi screening

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Involvement of 2'-5'oligoadenylate synthetase-like proteinin the survivalof Mycobacterium tuberculosis avirulent strain in macrophages

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Animal Diseases 2023年第4期3卷 275-285页

作者： Aikebaier Reheman Xiaojian Cao Yifan Wang Xi Nie Gang Cao Wei Zhou Bing Yang Yingying Lei Weipan Zhang muhammad ahsan naeem Xi Chen State Key Laboratory of Agricultural Microbiology College of Veterinary MedicineHuazhong Agricultural UniversityWuhan 430070China College of Animal Science and Technology Tarim UniversityAlar 843300China Bio-Medical Center Huazhong Agricultural UniversityWuhan 430070China Department of Basic Sciences(Pharmacology) University of Veterinary and Animal Sciences(Narowal Campus)Lahore 54000Pakistan

Mycobacterium tuberculosis(***)can replicate in the macrophage by interfering with many host protein *** it is far from known these host proteins for controlling *** ***,we infected macrophages including THP-1 and Raw264.7 cells with *** and identified the differentially expressed genes(DEGs)in the interferon signaling *** them,2'-5'oligoadenylate synthetase-like(OASL)underwent the greatest upregulation in ***-infected *** of the expression of OASL attenuated *** survival in ***,bioinformatics analysis revealed the potential interaction axis of OASL-TAB3-RvO127,which was further validated by the yeast-two-hybrid(Y2H)assay and *** interaction axis might regulate the *** survival and proliferation in *** study reveals a possible role of OASL during *** infection as a target to control its propagation.

关键词： Mycobacterium tuberculosis Interferon Interferon stimulated genes OASL

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Countrywide Survey for MERS-Coronavirus Antibodies in Dromedaries and Humans in Pakistan

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Virologica Sinica 2018年第5期33卷 410-417页

作者： Ali Zohaib muhammad Saqib muhammad Ammar Athar Jing Chen Awais-ur-Rahman Sial Saeed Khan Zeeshan Taj Halima Sadia Usman Tahir muhammad Haleem Tayyab muhammad Asif Qureshi muhammad Khalid Mansoor muhammad ahsan naeem Bing-Jie Hu Bilal Ahmed Khan Ikram Din Ujjan Bei Li Wei Zhang Yun Luo Yan Zhu Cecilia Waruhiu Iahtasham Khan Xing-Lou Yang muhammad Sohail Sajid Victor Max Corman Bing Yan Zheng-Li Shi CAS Key Laboratory of Special Pathogens and Biosafety Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China University of Ctunese Academy of Sciences Beijing 100049China Faculty of Veterinary Medicine University of AgricultureFaisalabad 38040Pakistan Laboratory of Cancer Biomarkers and Liquid Biopsy College of PharmacyHenan UniversityHenan 475004China Department of Clinical Studies Faculty of Veterinary and Animal SciencesPMAS-Arid Agriculture UniversityRawalpindi 46000Pakistan Dow University of Health Science Pakistan Karachi 74200 Department of Microbiology Government College University FaisalabadFaisalabad 38000Pakistan Livestock and Dairy Development Department(L&DD) Lahore 54000Pakistan College of Veterinary and Animal Sciences The Islamia University of BahawalpurBahawalpur 63100Pakistan State Key Laboratory of Agricultural Mlcroblology Huazhong Agricultural UniversityWuhan 430070China Liaqat University of Medical and Health Science Pakistan Jamshoro 76090 Section of Epidemiology and Public Health Department of Chnical SciencesUniversity of Veterinary and Animal SciencesLahore subcampusJhang 35200Pakistan Institute of Virology Chadte-Universitatsmedizin BerlinCorporate member of Free UniversityHumboldt-University and Berlin Institute of HealthBerlin 10117Germany German Center for Infection Research(DZIF) Berlin 10117Germany

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a zoonotic pathogen capable of causing severe respiratory disease in humans. Although dromedary camels are considered as a major reservoir host, the MERS-CoV infection dynamics in camels are not fully understood. Through surveillance in Pakistan, nasal (n = 776) and serum (n = 1050)samples were collected from camels between November 2015 and February 2018. Samples were collected from animal markets, free-roaming herds and abattoirs. An in-house ELISA was developed to detect IgG against MERS-CoV. A total of 794 camels were found seropositive for MERS-CoV. Prevalence increased with the age and the highest seroprevalence was recorded in camels aged [ 10 years (81.37%) followed by those aged 3.1–10 years (78.65%) and B 3 years (58.19%).Higher prevalence was observed in female (78.13%) as compared to male (70.70%). Of the camel nasal swabs, 22 were found to be positive by RT-qPCR though with high Ct values. Moreover, 2,409 human serum samples were also collected from four provinces of Pakistan during 2016–2017. Among the sampled population, 840 humans were camel *** we found a high rate of MERS-CoV antibody positive dromedaries (75.62%) in Pakistan, no neutralizing antibodies were detected in humans with and without contact to camels.

关键词： Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Camel -Human Pakistan

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