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A CrISPr knockout negative screen reveals synergy between CdKs inhibitor and metformin in the treatment of human cancer in vitro and in vivo

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Signal Transduction and Targeted Therapy 2020年第1期5卷 1091-1101页

作者： Yarui Ma Qing Zhu Junbo Liang Yifei Li Mo Li Ying Zhang Xiaobing Wang Yixin Zeng Yuchen Jiao State key laboratory of Molecular Oncology National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical College100021 BeijingChina State key laboratory of Medical Molecular Biology Institute of Basic Medical SciencesChinese Academy of Medical SciencesPeking Union Medical College10005 BeijingChina key laboratory of gene editing screening and r&d of digestive system tumor drugs Chinese Academy of Medical Sciences and Peking Union Medical College100021 BeijingChina State key laboratory of Oncology in South China Collaborative Innovation Center for Cancer MedicineSun Yat-sen University Cancer CenterGuangzhouChina department of Clinical laboratory National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical College100021 BeijingChina

laboratory research and pharmacoepidemiology provide support for metformin as a potential antitumor ***,the lack of a clear understanding of the indications of metformin limits its ***,we performed a genome-wide CrISPr knockout negative screen to identify potential targets that might synergize with ***-generation sequencing of pooled genomic dNAs isolated from surviving cells after 18 days of metformin treatment(T18)compared to those of the untreated cells at day 0(T0)yielded candidate *** of a group of cyclin-dependent kinases(CdKs),including CdK1,CdK4,and CdK6,confirmed the results of the *** treatment of the CdKs inhibitor abemaciclib with metformin profoundly inhibited tumor viability in vitro and in *** cell cycle parameters were not further altered under the combination treatment,investigation of the metabolome revealed significant changes in cell metabolism,especially with regard to fatty acid oxidation,the tricarboxylic acid cycle and aspartate *** changes appeared to be mediated through inhibition of the mTOr ***,our study suggests that the combination of CdKs inhibitor with metformin could be recognized as a potential therapy in future clinical applications.

关键词： epidemiology treatment CdKs

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AHr mediates the aflatoxin B1 toxicity associated with hepatocellular carcinoma

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Signal Transduction and Targeted Therapy 2021年第9期6卷 2809-2821页

作者： Qing Zhu Yarui Ma Junbo Liang Zhewen Wei Mo Li Ying Zhang Mei Liu Huan He Chunfeng Qu Jianqiang Cai Xiaobing Wang Yixin Zeng Yuchen Jiao State key laboratory of Molecular Oncology National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina State key laboratory of Medical Molecular Biology Institute of Basic Medical Sciences Chinese Academy of Medical SciencesSchool of Basic Medicine Peking Union Medical CollegeBeijingChina department of Hepatobiliary Surgery National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina key laboratory of gene editing screening and r&d of digestive system tumor drugs Chinese Academy of Medical SciencesPeking Union Medical CollegeBeijingChina State key laboratory of Oncology in South China Collaborative Innovation Center for Cancer MedicineSun Yat-sen University Cancer CenterGuangzhouChina department of Clinical laboratory National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina

Aflatoxin exposure is a crucial factor in promoting the development of primary hepatocellular carcinoma(HCC)in individuals infected with the hepatitis ***,the molecular pathways leading to its bioactivation and subsequent toxicity in hepatocytes have not been ***,we carried out a genome-wide CrISPr-Cas9 genetic screen to identify aflatoxin B1(AFB1)*** the most significant hits was the aryl hydrocarbon receptor(AHr),a ligand-binding transcription factor regulating cell metabolism,differentiation,and immunity.

关键词： aflatoxin metabolism hepatocellular

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Adjuvant camrelizumab plus apatinib in resected hepatocellular carcinoma with microvascular invasion:a multi-center real world study

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Hepatobiliary Surgery and Nutrition 2024年第4期13卷 616-631,I0022-I0024页

作者： Jingzhong Ouyang Yi Yang Yanzhao Zhou Xu Chang Zhengzheng Wang Qingjun Li Yu Tang Jianqiang Cai Jinxue Zhou Zhen Huang Hong Zhao department of Hepatobiliary and Pancreatic Surgery The Affiliated Cancer Hospital of Zhengzhou University&Henan Cancer HospitalZhengzhouChina department of Hepatobiliary Surgery National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina key laboratory of gene editing screening and research and development(r&d)of digestive system tumor drugs Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina department of Interventional Therapy II Shandong Cancer Hospital and InstituteShandong First Medical University and Shandong Academy of Medical SciencesJinanChina Clinical Trials Center National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina

Background:It is well known that laparoscopic liver surgery can offer advantages over open liver surgery in selected ***,what type of procedures can benefit most from a laparoscopic approach has been investigated poorly thus *** aim of this study is thus to define the extent of advantages of laparoscopic over open liver surgery for lesions in the anterolateral(AL)and posterosuperior(PS)***:In this international multicentre retrospective cohort study,laparoscopic and open minor liver resections for lesions in the AL and PS segments were compared after propensity score *** differential benefit of laparoscopy over open liver surgery,calculated using bootstrap sampling,was compared between AL and PS resections and expressed as a delta of the ***:After matching,3,040 AL and 2,336 PS resections were compared,encompassing open and laparoscopic procedures in a 1:1 *** and PS laparoscopic liver resections were more advantageous in comparison to open in terms of blood loss,transfusion rate,complications,and length of ***,AL resections benefitted more from laparoscopy than PS in terms of overall and severe complications(d-difference were 4.8%,P=0.046 and 3%,P=0.046)and blood loss(d-difference was 195 mL,Pr results were observed in the subset for high-volume centres,while in recent years no significant differences were found in the differential benefit between AL and PS ***:The advantage of laparoscopic over open liver surgery is greater in the AL segments than in the PS segments.

关键词： Liver neoplasms hepatectomy laparoscopic liver resection open liver resection

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Novel gold nanoparticles targeting somatostatin receptor subtype two with near-infrared light for neuroendocrine tumour therapy

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Nano research 2022年第10期15卷 9149-9159页

作者： Qichen Chen Zilin Li Jiangyuan Yu Qing Xie Haizhen Lu Yiqiao deng Jinghua Chen Wenjia Zhu Li Huo Yizhou Zhang Wei Song Jianqiang Lan Jianqiang Cai Zhen Huang Zixi Wang Hong Zhao department of Hepatobiliary Surgery State Key Laboratory of Molecular OncologyNational Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100021China key laboratory of gene editing screening and r&d of digestive system tumor drugs Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100021China State key laboratory of Tribology Department of Mechanical EngineeringTsinghua UniversityBeijing 100084China department of Nuclear Medicine Key Laboratory of Carcinogenesis and Translational ResearchMinistry of EducationPeking University Cancer Hospital&InstituteBeijing 100142China department of Pathology National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeNo.17Panjiayuan NanliBeijing 100021China Nuclear Medicine department State Key Laboratory of Complex Severe and Rare DiseasesCenter for Rare Diseases ResearchBeijing Key Laboratory of Molecular Targeted Diagnosis and Therapy in Nuclear MedicinePeking Union Medical College HospitalChinese Academy of Medical Science and Peking Union Medical CollegeBeijing 100005China department of Biochemistry and Molecular Biology State Key Laboratory of Medical Molecular BiologyInstitute of Basic Medical Sciences Chinese Academy of Medical SciencesSchool of Basic MedicinePeking Union Medical CollegeBeijing 100005China Guangdong research Center of Organoid Engineering and Technology Accurate International Biotechnology Co.Ltd.Guangzhou 510650China

Neuroendocrine tumours(NETs)are rare cancers with positive somatostatin receptor 2(SSTr2)expression,and treatment strategies for NETs are not ***-mediated therapy targeting SSTr2 in NETs is very *** study firstly combined mesoporous silica-coated gold nanorods(AuNrs@mSiO_(2))and targeting-SSTr2 dodecane tetraacetic acidtyrosine3-octreotate(dOTA-TATE)into AuNrs@mSiO_(2)@dOTA-TATE to investigate NETs inhibition under near-infrared ***@mSiO_(2)@dOTA-TATE showed good photothermal conversion *** vitro,under light irradiation,the cell viability significantly decreased with increasing AuNr@mSiO_(2)@dOTA-TATE concentration;in two successfully established neuroendocrine tumour organoids with SSTr2 expression,AuNrs@mSiO_(2)@dOTA-TATE with light inhibited tumours significantly better than AuNrs@mSiO_(2) with *** vivo,the SSTr2-targeting ability and biodistribution of AuNrs@mSiO_(2)@dOTA-TATE were confirmed with AuNrs@mSiO_(2)@64Cu-dOTA-TATE under micro-positron emission tomography/computed tomography(micro-PET/CT);in the AuNrs@mSiO_(2)@dOTA-TATE with laser group,the tumour surface temperature increased rapidly,with tumour volumes similar to those in the octreotide group and significantly lower than those in other *** was no significant difference in mice body weight between the AuNrs@mSiO_(2)@dOTA-TATE with laser group and other *** significant inflammatory lesions or cell necrosis was found in the main *** summary,we presented a feasible strategy to construct AuNrs@mSiO_(2)@dOTA-TATE with good photothermal conversion efficiency,targetingSSTr2 ability,significant antitumour effects,and good biocompatibility,warranting further explorations of AuNrs@mSiO_(2)@dOTA-TATE for NETs therapy applications.

关键词： neuroendocrine tumours somatostatin receptor 2 nanoparticles photothermal therapy

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rING finger 138 deregulation distorts NF-κB signaling and facilities colitis switch to aggressive malignancy

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Signal Transduction and Targeted Therapy 2022年第7期7卷 2555-2567页

作者： Yalan Lu rong Huang Jianming Ying Xingchen Li Tao Jiao Lei Guo Haitao Zhou Han Wang Amannisa Tuersuntuoheti Jianmei Liu Qichen Chen Yanhong Wang Luying Su Changyuan Guo Fu Xu Ziyi Wang Yan Lu Kai Li Junbo Liang Zhen Huang Xiao Chen Jinjie Yao Hanjie Hu Xiaowen Cheng Yufeng Wan Xinyan Chen Ning Zhang Shiying Miao Jianqiang Cai Linfang Wang Changzheng Liu Wei Song Hong Zhao department of Biochemistry and Molecular Biology State Key Laboratory of Medical Molecular BiologyInstitute of Basic Medical Sciences Chinese Academy of Medical SciencesSchool of Basic Medicine Peking Union Medical CollegeBeijing 100005China department of Hepatobiliary Surgery State Key Laboratory of Molecular OncologyNational Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100021China key laboratory of gene editing screening and r&d of digestive system tumor drugs Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100021China key laboratory of Human disease Comparative Medicine Chinese Ministry of HealthBeijing Key Laboratory for Animal Models of Emerging and Remerging Infectious DiseasesInstitute of Laboratory Animal ScienceChinese Academy of Medical Sciences and Comparative Medicine CenterPeking Union Medical CollegeBeijing 100021China National Cancer Center/National Clinical research Center for Cancer/Cancer Hospital&Shenzhen Hospital Chinese Academy of Medical Sciences and Peking Union Medical CollegeShenzhen 518116China department of Pathology State Key Laboratory of Molecular OncologyNational Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100021China department of Clinical laboratory the First Affiliated HospitalAnhui Medical UniversityHefei 230022China Wellcome Centre for Anti-Infectives research(WCAIr) Division of Biological Chemistry and Drug DiscoverySchool of Life SciencesUniversity of DundeeDundee DD15EHUK

Prolonged activation of nuclear factor(NF)-кB signaling significantly contributes to the development of colorectal cancer(CrC).New therapeutic opportunities are emerging from targeting this distorted cell signaling ***,we discovered the critical role of rING finger 138(rNF138)in CrC tumorigenesis through regulating the NF-кB signaling,which is independent of its Ubiquitin-E3 ligase activity involved in dNA damage ***138^(−/−) mice were hyper-susceptible to the switch from colitis to aggressive malignancy,which coincided with sustained aberrant NF-кB signaling in the colonic ***,rNF138 suppresses the activation of NF-кB signaling pathway through preventing the translocation of NIK and IKK-Beta Binding Protein(NIBP)to the cytoplasm,which requires the ubiquitin interaction motif(UIM)*** importantly,we uncovered a significant correlation between poor prognosis and the downregulation of rNF138 associated with reinforced NF-кB signaling in clinical settings,raising the possibility of rNF138 dysregulation as an indicator for the therapeutic intervention targeting NF-кB *** the xenograft models built upon either rNF138-dificient CrC cells or the cells derived from the rNF138-dysregulated CrC patients,we demonstrated that the inhibition of NF-кB signaling effectively hampered tumor ***,our work defined the pathogenic role of aberrant NF-кB signaling due to rNF138 downregulation in the cascade events from the colitis switch to colonic neoplastic transformation and progression,and also highlights the possibility of targeting the NF-кB signaling in treating specific subtypes of CrC indicated by rNF138-ablation.

关键词： malignancy sustained finger

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Correction: AHr mediates the Aflatoxin B1 toxicity associated with hepatocellular carcinoma

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Signal Transduction and Targeted Therapy 2022年第1期7卷 331-332页

作者： Qing Zhu Yarui Ma Junbo Liang Zhewen Wei Mo Li Ying Zhang Mei Liu Huan He Chunfeng Qu Jianqiang Cai Xiaobing Wang Yixin Zeng Yuchen Jiao State key laboratory of Molecular Oncology National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China key laboratory of gene editing screening and r&d of digestive system tumor drugs Chinese Academy of Medical Sciences Peking Union Medical College Beijing China State key laboratory of Oncology in South China Collaborative Innovation Center for Cancer Medicine Sun Yat-sen University Cancer Center Guangzhou China department of Clinical laboratory National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China State key laboratory of Medical Molecular Biology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences School of Basic Medicine Peking Union Medical College Beijing China department of Hepatobiliary Surgery National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China

Correction to:Signal Transduction and Targeted Therapy https://***/10.1038/s41392-021-00713-1 z published online 9 August 2021 After online publication of the article^(1),the authors noticed one inadvertent mistake occurred during the production process in Fig.7 that needs to be *** correct data are provided as *** key findings of the article are not affected by these *** original article has been corrected.

关键词： correction finding corrected

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Cd47KO/CrT dual-bioengineered cell membrane-coated nanovaccine combined with anti-Pd-L1 antibody for boosting tumor immunotherapy

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Bioactive Materials 2023年第4期22卷 211-224页

作者： Shengyang Liu Jiayan Wu Yuanji Feng Xiaoya Guo Tong Li Meng Meng Jie Chen daquan Chen Huayu Tian School of Pharmacy Key Laboratory of Molecular Pharmacology and Drug Evaluation(Yantai University)Ministry of EducationCollaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of ShandongYantai UniversityYantai264005PR China key laboratory of Polymer Ecomaterials Changchun Institute of Applied ChemistryChinese Academy of SciencesChangchun130022PR China College of Chemistry and Chemical Engineering Xiamen UniversityXiamen361005PR China

tumor vaccines trigger tumor-specific immune responses to prevent or treat tumors by activating the hosts’immune systems,and therefore,these vaccines have potential clinical ***,the low immunogenicity of the tumor antigen itself and the low efficiency of the vaccine delivery system hinder the efficacy of tumor vaccines that cannot produce high-efficiency and long-lasting antitumor immune ***,we constructed a nanovaccine by integrating Cd47KO/CrT dual-bioengineered B16F10 cancer cell membranes and the unmethylated cytosine-phosphate-guanine(CpG)*** PEI25k was used to load unmethylated cytosine-phosphate-guanine(CpG)through electrostatic adsorption to prepare PEI25k/CpG nanoparticles(PEI25k/CpG-NPs).Cd47KO/CrT dual-bioengineered cells were obtained by CrISPr-Cas9 gene editing technology,followed by the cell surface translocation of calreticulin(CrT)to induce immunogenic cell death(ICd)in ***,the extracted cell membranes were coextruded with PEI25k/CpG-NPs to construct the Cd47KO/CrT dual-bioengineered cancer cell membrane-coated nanoparticles(dBE@CCNPs).dBE@CCNPs could promote endocytosis of antigens and adjuvants in murine bone marrow derived dendritic cells(BMdCs)and induce their maturation and antigen *** avoid immune checkpoint molecule-induced T cell dysfunction,the immune checkpoint inhibitor,the anti-Pd-L1 antibody,was introduced to boost tumor immunotherapy through a combination with the dBE@CCNPs *** combination therapy strategy can significantly alleviate tumor growth and may open up a potential strategy for clinical tumor immunotherapy.

关键词： CrISPr-Cas9 dual-bioengineered cell membrane Immune checkpoint blockade Nanovaccine tumor immunotherapy

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Single-cell exome sequencing reveals polyclonal seeding and TrPS1 mutations in colon cancer metastasis

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Signal Transduction and Targeted Therapy 2024年第10期9卷 4637-4652页

作者： Jianqiang Cai Weilong Zhang Yalan Lu Wenjie Liu Haitao Zhou Mei Liu Xinyu Bi Jianmei Liu Jinghua Chen Yanjiang Yin Yiqiao deng Zhiwen Luo Yi Yang Qichen Chen Xiao Chen Zheng Xu Yueyang Zhang Chaoling Wu Qizhao Long Chunyuan Huang Changjian Yan Yan Liu Lei Guo Weihua Li Pei Yuan Yucheng Jiao Wei Song Xiaobing Wang Zhen Huang Jianming Ying Hong Zhao department of Hepatobiliary Surgery State Key Laboratory of Molecular OncologyNational Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina key laboratory of gene editing screening and r&d of digestive system tumor drugs Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina department of Hematology Lymphoma Research CenterPeking University Third HospitalBeijingChina key laboratory of Human disease Comparative Medicine Chinese Ministry of HealthBeijing Key Laboratory for Animal Models of Emerging and Remerging Infectious DiseasesInstitute of Laboratory Animal ScienceChinese Academy of Medical Sciences and Comparative Medicine CenterPeking Union Medical CollegeBeijingChina department of Biochemistry and Molecular Biology State Key Laboratory of Medical Molecular BiologyInstitute of Basic Medical Sciences Chinese Academy of Medical SciencesSchool of Basic Medicine Peking Union Medical CollegeBeijingChina department of Colorectal Surgery State Key Laboratory of Molecular OncologyNational Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina State key laboratory of Molecular Oncology National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina department of Pathology State Key Laboratory of Molecular OncologyNational Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina

Liver metastasis remains the primary cause of mortality in patients with colon cancer. Identifying specific driver gene mutations that contribute to metastasis may offer viable therapeutic targets. To explore clonal evolution and genetic heterogeneity within the metastasis, we conducted single-cell exome sequencing on 150 single cells isolated from the primary tumor, liver metastasis, and lymphatic metastasis from a stage IV colon cancer patient. The genetic landscape of the tumor samples revealed that both lymphatic and liver metastases originated from the same region of the primary tumor. Notably, the liver metastasis was derived directly from the primary tumor, bypassing the lymph nodes. Comparative analysis of the sequencing data for individual cell pairs within different tumors demonstrated that the genetic heterogeneity of both liver and lymphatic metastases was also greater than that of the primary tumor. This finding indicates that liver and lymphatic metastases arose from clusters of circulating tumor cell (CTC) of a polyclonal origin, rather than from a single cell from the primary tumor. Single-cell transcriptome analysis suggested that higher EMT score and CNV scores were associated with more polyclonal metastasis. Additionally, a mutation in the TrPS1 (Transcriptional repressor GATA binding 1) gene, TrPS1 r544Q, was enriched in the single cells from the liver metastasis. The mutation significantly increased CrC invasion and migration both in vitro and in vivo through the TrPS1r544Q/ZEB1 axis. Further TrPS1 mutations were detected in additional colon cancer cases, correlating with advanced-stage disease and inferior prognosis. These results reveal polyclonal seeding and TrPS1 mutation as potential mechanisms driving the development of liver metastases in colon cancer.

关键词： metastasis colon cancer

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Logic-gated tumor-microenvironment nanoamplifier enables targeted delivery of CrISPr/Cas9 for multimodal cancer therapy

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Acta Pharmaceutica Sinica B 2024年第2期14卷 795-807页

作者： Yongchun Pan Xiaowei Luan Fei Zeng Xuyuan Wang Shurong Qin Qianglan Lu Guanzhong He Yanfeng Gao Xiaolian Sun Xin Han Bangshun He Yujun Song department of laboratory Medicine Nanjing First HospitalNanjing Medical UniversityNanjing 210006China College of Engineering and Applied Sciences State Key Laboratory of Analytical Chemistry for Life ScienceNanjing UniversityNanjing 210023China State key laboratory of Natural Medicines Key Laboratory of Drug Quality Control and PharmacovigilanceDepartment of Pharmaceutical AnalysisChina Pharmaceutical UniversityNanjing 211198China School of Medicine&Holistic Integrative Medicine Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources IndustrializationNanjing University of Chinese MedicineNanjing 210023China

recent innovations in nanomaterials inspire abundant novel tumor-targeting CrISPr-based gene ***,the therapeutic efficiency of traditional targeted nanotherapeutic strategies is limited by that the biomarkers vary in a spatiotemporal-dependent manner with tumor ***,we propose a self-amplifying logic-gated gene editing strategy for gene/H_(2)O_(2)-mediated/starvation multimodal cancer *** this approach,a hypoxia-degradable covalent-organic framework(COF) is synthesized to coat a-ZIF-8 in which glucose oxidase(GOx) and CrISPr system are *** intensify intracellular redox dyshomeostasis,dNAzymes which can cleave catalase mrNA are loaded as *** the nano system gets into the tumor,the weakly acidic and hypoxic microenvironment degrades the ZIF-8@COF to activate GOx,which amplifies intracellular H^(+)and hypoxia,accelerating the nanocarrier degradation to guarantee available CrISPr plasmid and GOx release in target *** tandem reactions deplete glucose and oxygen,leading to logic-gated-triggered gene editing as well as synergistic gene/H_(2)O_(2)-mediated/starvation ***,this approach highlights the biocomputing-based CrISPr delivery and underscores the great potential of precise cancer therapy.

关键词： CrISPrdelivery Enzymeencapsulation Logic gate Hybridmaterial Hypoxia gene editing Precise nanomedicine Multimodal therapy

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reprogramming Mycobacterium tuberculosis CrISPr system for gene editing and Genomewide rNA Interference screening

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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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