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Large-Scale CRISPR Screen of LDLR Pathogenic Variants

Research 2024年第2期2023卷 213-225页

作者： Mengjing Li Lerong Ma Yiwu Chen Jianing Li Yanbing Wang Wenni You Hongming Yuan Xiaochun Tang Hongsheng Ouyang Daxin Pang Key Lab for Zoonoses Research Ministry of EducationAnimal Genome Editing Technology Innovation CenterCollege of Animal SciencesJilin UniversityChangchunJilin Province 130062China The Institute of Translational Medicine Tianjin Union Medical Center of Nankai UniversityTianjin 300071China Chongqing researchinstitute Jilin UniversityChongqing 401123China Chongqing Jitang biotechnology Research Institute Co.Ltd. ChongqingChina

Familial hypercholesterolemia(FH)is a frequently occurring genetic disorder that is linked to early-onset cardiovascular *** left untreated,patients with this condition can develop severe cardiovascular ***,many patients remain undiagnosed,and even when diagnosed,the treatment is often not *** mutations in the LDLR gene are the primary cause of FH,predicting whether novel variants are pathogenic is not a straightforward *** the functionality of LDLR variants is crucial in uncovering the genetic basis of *** study utilized CRISPR/Cas9 cytosine base editors in pooled screens to establish a novel approach for functionally assessing tens of thousands of LDLR variants on a large scale.A total of more than 100 single guide RNAs(sgRNAs)targeting LDLR pathogenic mutations were successfully screened with relatively high *** of these,5 sgRNAs were further subjected to functional verification studies,including 1 in the promoter,1 in the antisense RNA,1 in the exon,and 2 in the *** for the variant caused by the sgRNA located at intron 16,the functionalities of the other LDLR variants were all *** high similarity of LDLR intron sequences may lead to some false ***,these results confirm the reliability of the large-scale screening strategy for functional analysis of LDLR variants,and the screened candidate pathogenic mutations could be used as an auxiliary means of clinical gene detection to prevent FH-induced heart disease.

关键词： LDLR Screen straight

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Identification of squamous cell carcinoma associated proteins by proteomics and loss of beta tropomyosin expression in esophageal cancer

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World Journal of Gastroenterology 2006年第44期12卷 7104-7112页

作者： Ferdous Rastgar Jazii Zahra Najafi Reza Malekzadeh Thomas P Conrads Abed Ali Ziaee Christian Abnet Mansour Yazdznbod Ali Asghar Karkhane Ghasem H Salekdeh National Institute of Genetic Engineering and biotechnology POBOX: 14155 6343 Tehran Iran Digestive Disease Research Center Shariati Hospital Medical University Tehran Iran Laboratory of Proteomics and Analytical Technologies SAIC -Frederick B Frederick MD 21702 United States Institute of Biochemistry and Biophysics Tehran University Tehran Iran NCI Center for Cancer Research CancerPrevention Studies Branch Bethesda United States Department of surgery Shariati Hospital Medical University Tehran Iran agricultural biotechnology researchinstitute of iran Karaj Iran

AIM： To assess the proteome of normal versus tumor tissue in squamous cell carcinoma of the esophagus （SCCE） in iranian patients and compare our results with former reports by using proteomics. METHODS： Protein was extracted from normal and tumor tissues. Two dimensional electrophoresis was carried out and spots with differential expression were identified with mass spectrometry. RNA extraction and RT-PCR along with immunodetection were performed. RESULTS： Fourteen proteins were found whose expression levels differed in tumor compared to normal tissues. Mass spectrometric analysis resulted in the identification of β-tropomyosin （TMβ）, myosin light chain 2 （and its isoform）, myosin regulatory light chain 2, peroxyredoxin 2, annexin I and an unknown polypeptide as the down regulated polypeptides in tumor tissue. Heat shock protein 70 （HSP70）, TPM4-ALK fusion oncoprotein 2, myosin light polypeptide 6, keratin I, GH16431p and calreticulin were the up-regulated polypeptides found in tumor tissue. Several of these proteins, such as TMβ, HSP70, annexin Ⅰ, calreticulin, TPM4-ALK and isoforms of myosins, have been well recognized in tumorigenesis of esophageal or other types of cancers. CONCLUSION： Our study not only supports the involvement of some of the formerly reported proteins in SCCE but also introduces additional proteins found to be lost in SCCE, including TMβ.

关键词： Squamous Cell Carcinoma Esophagus Esophageal Proteomics Two dimensional electrophoresis Polypeptide marker

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三种杀虫剂胁迫下西花蓟马(Frankliniella occidentalis)共响应基因的鉴定与分析

三种杀虫剂胁迫下西花蓟马(Frankliniella occidentalis)共响应基...

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中国植物保护学会2019年学术年会

作者：高岳 Min Ju Kim In Hong Jeong J.Marshall Clark Si Hyeock Lee Department of agricultural biotechnology Seoul National University Division of Crop Protection National Insstitute of Agricultural ScienceRural Development Administration Department of Veterinary & Animal Sciences University of Massachusetts at Amherst researchinstitute for Agriculture and Life Science Seoul National University

西花蓟马(Frankliniella occidentalis)是一种重大的园艺害虫,在世界多个国家发生危害,造成了巨大的经济损失。由于杀虫剂施用仍是防治西花蓟马的主要措施,现在其已经对多种杀虫剂产生了较高水平的抗性,进一步增加了其防治的难度。为实... 详细信息

西花蓟马(Frankliniella occidentalis)是一种重大的园艺害虫,在世界多个国家发生危害,造成了巨大的经济损失。由于杀虫剂施用仍是防治西花蓟马的主要措施,现在其已经对多种杀虫剂产生了较高水平的抗性,进一步增加了其防治的难度。为实现对西花蓟马种群的合理控制,开展对西花蓟马抗性机制的研究成为必要举措之一。基于此,本研究选用虫螨腈、呋虫胺和多杀菌素3种不同类型杀虫剂,运用RCVpW法(Residual Contact Vial plus Water)以亚致死剂量对西花蓟马进行处理,并进行数字表达谱分析。结果表明,3种杀虫剂亚致死剂量胁迫下,西花蓟马体内共有230个基因发生共响应,其中199个上调,31个下调。GO分析结果显示,不同杀虫剂胁迫下,西花蓟马GO通路分布情况不同;大量共上调的基因同蛋白质、脂质和碳水化合物代谢相关;同时筛选出10个解毒代谢酶基因,包括4个细胞色素P450基因(Cyp6a1、Cyp6a13、Cyp6a14、Cyp6k1)、4个UGT基因(UGT2B2、UGT2B15、UGT2B17、UGT2C1)、1个谷胱甘肽S转移酶基因和1个ATP结合盒转运体基因(ABC transporter G20)。此外,笔者发现3种杀虫剂胁迫下,3个含有CH-型锌指结构的转录调节基因mRNA表达水平均下调。上述研究结果暗示了上述共响应基因可能介入了西花蓟马的通用抗性机制,但其具体的角色和机理则需要进一步的研究。

关键词：西花蓟马杀虫剂胁迫亚致死剂量数字表达谱细胞色素P450

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Remodeling of the plasma for sperm-egg recognition： proteins membrane in preparation roles of acrosomal

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Asian Journal of Andrology 2015年第4期17卷 574-582页

作者： Nongnuj Tanphaichitr Kessiri Kongmanas Hathairat Kruevaisayawan Arpornrad Saewu Clarissa Sugeng Jason Fernandes Puneet Souda Jonathan B Angel Kym F Faull R John Aitken Julian Whitelegge Daniel Hardy Trish Berger Mark Baker Chronic Disease Program Ottawa Hospital ResearchInstitute Ottawa Ontario Canada Department ofObstetrics and Gynaecology University of OttawaOntario Canada Department of Biochemistry Microbiology and Immunology University of OttawaOntario Canada Department of Anatomy Faculty ofMedical Sciences Naresuan University PhitsanulokThailand Division of Infectious Diseases OttawaHospital - General Campus Ottawa Ontario Canada Pasarow Mass Spectrometry Laboratory Universityof California Los Angeles California USA TheARC Centre of Excellence in biotechnology andDevelopment School of Environmental and LifeSciences University of Newcastle Callaghan NewSouth Wales Australia Department of Cell Biologyand Biochemistry Health Sciences Center Texas TechUniversity Texas USA Department of Animal ScienceUniversity of California Davis California USA

The interaction of sperm with the egg＇s extracellular matrix, the zona pellucida （ZP） is the first step of the union between male and female gametes. The molecular mechanisms of this process have been studied for the past six decades with the results obtained being both interesting and confusing. In this article, we describe our recent work, which attempts to address two lines of questions from previous studies. First, because there are numerous ZP binding proteins reported byvarious researchers, how do these proteins act together in sperm-ZP interaction？ Second, why do a number of acrosomal proteins have ZP affinity？ Are they involved mainly in the initial sperm-ZP binding or rather in anchoring acrosome reacting/reacted spermatozoa to the ZP？ Our studies reveal that a number of ZP binding proteins and chaperones, extracted from the anterior sperm head plasma membrane, coexist as high molecular weight （HMW） complexes, and that these complexes in capacitated spermatozoa have preferential ability to bind to the ZP. Zonadhesin （ZAN）, known as an acrosomal protein with ZP affinity, is one of these proteins in the HMW complexes. Immunoprecipitation indicates that ZAN interacts with other acrosomal proteins, proacrosin/acrosin and sp32 （ACRBP）, also present in the HMW complexes. Immunodetection of ZAN and proacrosin/acrosin on spermatozoa further indicates that both proteins traffic to the sperm head surface during capacitation where the sperm acrosomal matrix is still intact, and therefore they are likely involved in the initial sperm-ZP binding step.

关键词： SPERM CAPACITATION ANDSPERM-ZONA PELLUCIDAINTERACTION- BACKGROUND ANDCONFUSION IN THE FIELDS

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Characterizing the tumor microenvironment at the single-cell level reveals a novel immune evasion mechanism in osteosarcoma

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Bone Research 2023年第1期11卷 124-135页

作者： Weijian Liu Hongzhi Hu Zengwu Shao Xiao Lv Zhicai Zhang Xiangtian Deng Qingcheng Song Yong Han Tao Guo Liming Xiong Baichuan Wang Yingze Zhang Department of Orthopaedics Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430022China Department of Orthopaedic Surgery The Third Hospital of Hebei Medical UniversityShijiazhuang050051China Orthopaedic Institute of Hebei Province Shijiazhuang050051China Key Laboratory of Biomechanics of Hebei Province Shijiazhuang050051China Animal Center of Hebei Ex&In vivo biotechnology Shijiazhuang050051China Department of Pharmacy Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina Department of Pathology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina

The immune microenvironment extensively participates in tumorigenesis as well as progression in osteosarcoma(OS).However,the landscape and dynamics of immune cells in OS are poorly *** analyzing single-cell RNA sequencing(sc RNA-seq)data,which characterize the transcription state at single-cell resolution,we produced an atlas of the immune microenvironment in *** results suggested that a cluster of regulatory dendritic cells(DCs)might shape the immunosuppressive microenvironment in OS by recruiting regulatory T *** also found that major histocompatibility complex class I(MHC-I)molecules were downregulated in cancer *** findings indicated a reduction in tumor immunogenicity in OS,which can be a potential mechanism of tumor immune *** note,CD24 was identified as a novel“don’t eat me”signal that contributed to the immune evasion of OS ***,our findings provide insights into the immune landscape of OS,suggesting that myeloid-targeted immunotherapy could be a promising approach to treat OS.

关键词： osteosarcoma microenvironment CD24

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