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Designing and generating a mouse model:frequently asked questions

The Journal of Biomedical Research 2021年第2期35卷 76-90页

作者： Channabasavaiah BGurumurthy Thomas LSaunders Masato Ohtsuka Department of Pharmacology and Experimental Neuroscience Department of Pharmacology and Experimental NeuroscienceCollege of MedicineUniversity of Nebraska Medical CenterOmahaNE 68106-5915USA Mouse Genome Engineering core facility Department of Pharmacology and Experimental NeuroscienceCollege of MedicineUniversity of Nebraska Medical CenterOmahaNE 68106-5915USA Department of Internal medicine Division of Genetic MedicineUniversity of Michigan Medical SchoolAnn ArborMI 48109USA Transgenic Animal Model core University of MichiganAnn ArborMI 48109USA Department of molecular Life Science Division of Basic Medical Science and Molecular MedicineTokai University School of MedicineIseharaKanagawa 259-1193Japan The Institute of Medical Sciences Tokai UniversityIseharaKanagawa 259-1193Japan

Genetically engineered mouse(GEM)models are commonly used in biomedical *** GEMs involve complex set of experimental procedures requiring sophisticated equipment and highly skilled technical *** of these reasons,most research institutes set up centralized core facilities where custom GEMs are created for research ***,on the other hand,when they begin thinking about generating GEMs for their research,several questions arise in their *** example,what type of model(s)would be best useful for my research,how do I design them,what are the latest technologies and tools available for developing my model(s),and finally how to breed GEMs in my *** there are several considerations and options in mouse designs,and as it is an expensive and time-consuming endeavor,careful planning upfront can ensure the highest chance of *** this article,we provide brief answers to several frequently asked questions that arise when researchers begin thinking about generating mouse model(s)for their work.

关键词： CRISPR transgenic mouse genetic engineering knockout mouse conditional knockout mouse knock-in mouse

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Newly developed Fe3O4–Cr2O3 magnetic nanocomposite for photocatalytic decomposition of 4-chlorophenol in water

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Journal of Environmental Sciences 2017年第2期29卷 333-340页

作者： Khoirakpam Kesho Singh Kula Kamal Senapati Chandan Borgohain Kanak Chandra Sarma Department of Instrumentation and USIC Gauhati UniversityGuwahati 781014AssamIndia Central Instruments facility Indian Institute of Technology GuwahatiGuwahati 781039AssamIndia Department of biochemistry Tocklai Tea Research InstituteTRAJorhat 785008AssamIndia

Chlorophenols, typically 4-chlorophenols are highly toxic and non-biodegradable organic contaminants which pose serious threat to the environment, particularly when released into aqueous medium. The removal of these pollutants by efficient method has received worldwide concern in recent past. A new Fe3O4–Cr2O3 magnetic nanocomposite was synthesized by wet chemical method under ultrasonic irradiation. Microstructure and morphology of the nanocomposite were characterized by powder X-ray diffraction（XRD）,Fourier transform infrared（FT-IR）, and a transmission electron microscope（TEM）. Magnetic and optical properties were studied by a vibrating sample magnetometer（VSM） and an ultraviolet–visible（UV–Vis） spectrophotometer respectively. The magnetic nanocomposite（MNC） was used as photocatalyst for effective decomposition of 4-chlorophenol in water under ultraviolet（UV） irradiation.

关键词： Chlorophenol Magnetic nanocomposite Photocatalyst core/shell nanocomposite

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Cytokine- and chemokine-induced inflammatory colorectal tumor microenvironment: Emerging avenue for targeted therapy

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Cancer Communications 2022年第8期42卷 689-715页

作者： Ajaz A.Bhat Sabah Nisar Mayank Singh Bazella Ashraf Tariq Masoodi Chandra P.Prasad Atul Sharma Selma Maacha Thasni Karedath Sheema Hashem Syed Besina Yasin Puneet Bagga Ravinder Reddy Michael P.Frennaux Shahab Uddin Punita Dhawan Mohammad Haris Muzafar A.Macha Laboratory of molecular and Metabolic Imaging Cancer Research DepartmentSidra MedicineDoha 26999Qatar Department of Medical Oncology Dr.B.R.Ambedkar Institute Rotary Cancer HospitalAll India Institute of Medical Sciences(AIIMS)New Delhi 110029India Department of Biotechnology School of Life SciencesCentral University of KashmirGanderbalJammu&Kashmir 191201India Division of Translational medicine Research BranchSidra MedicineDoha 26999Qatar Genomics core facility QBRIQatar FoundationDoha 34110Qatar Department of Pathology Sher-I-Kashmir Institute of Medical SciencesSrinagarJammu&Kashmir 190011India Department of Diagnostic Imaging St.Jude Children’s Research HospitalMemphisTN 38105USA Center for Advanced Metabolic Imaging in Precision medicine Department of RadiologyPerelman School of Medicine at the University of PennsylvaniaPhiladelphiaPA 19104USA Academic Health System Hamad Medical CorporationDoha 3050Qatar Translational Research Institute Hamad Medical CorporationDoha 3050Qatar Department of biochemistry and molecular Biology University of Nebraska Medical CenterOmahaNE 68198USA Laboratory Animal Research Center Qatar UniversityDoha 2713Qatar Watson-Crick Centre for molecular medicine Islamic University of Science and TechnologyAwantiporaJammu&Kashmir 192122India

Colorectal cancer(CRC)is a predominant life-threatening cancer,with liver and peritoneal metastases as the primary causes of *** inflammation,a known CRC risk factor,nurtures a local inflammatory environment enriched with tumor cells,endothelial cells,immune cells,cancer-associated fibroblasts,immunosuppressive cells,and secretory growth *** complex interactions of aberrantly expressed cytokines,chemokines,growth factors,and matrix-remodeling enzymes promote CRC pathogenesis and evoke systemic responses that affect disease *** evidence suggests that these cytokines and chemokines play a role in the progression of CRC through immunosuppression and modulation of the tumor microenvironment,which is partly achieved by the recruitment of immunosuppressive *** cells impart features such as cancer stem cell-like properties,drug resistance,invasion,and formation of the premetastatic niche in distant organs,promoting metastasis and aggressive CRC growth.A deeper understanding of the cytokineand chemokine-mediated signaling networks that link tumor progression and metastasis will provide insights into the mechanistic details of disease aggressiveness and facilitate the development of novel therapeutics for ***,we summarized the current knowledge of cytokine-and chemokine-mediated crosstalk in the inflammatory tumor microenvironment,which drives immunosuppression,resistance to therapeutics,and metastasis during CRC *** also outlined the potential of this crosstalk as a novel therapeutic target for *** major cytokine/chemokine pathways involved in cancer immunotherapy are also discussed in this review.

关键词： chemokine colorectal cancer cytokine drug resistance epithelial-mesenchymal transition immunosuppression immunotherapy inflammation metastasis tumor microenvironment

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CD137 agonist potentiates the abscopal efficacy of nanoparticle-based photothermal therapy for melanoma

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Nano Research 2022年第3期15卷 2300-2314页

作者： Preethi Bala Balakrishnan Debbie KLedezma Juliana Cano-Mejia Jaclyn Andricovich Erica Palmer Vishal APatel Patricia SLatham Eric SYvon Alejandro Villagra Rohan Fernandes Elizabeth E.Sweeney GW Cancer Center Department of MedicineSchool of Medicine and Health SciencesGeorge Washington UniversityWashingtonDC 20052USA The Institute for Biomedical Sciences School of Medicine and Health SciencesGeorge Washington UniversityWashingtonDC 20052USA GW Cancer Center Department of Biochemistry and Molecular MedicineSchool of Medicine and Health SciencesGeorge Washington UniversityWashingtonDC 20052USA Department of Dermatology&Oncology School of Medicine and Health SciencesGeorge Washington UniversityWashingtonDC 20037USA Department of Pathology School of Medicine and Health SciencesGeorge Washington UniversityWashingtonDC 20037USA ImmunoBlue BethesdaMD 20817USA

Despite the promise of immunotherapy such as the immune checkpoint inhibitors(ICIs)anti-PD-1 and anti-CTLA-4 for advanced melanoma,only 26%–52%of patients respond,and many experience grade III/IV immune-related adverse *** by the need for an effective therapy for patients non-responsive to clinically approved ICIs,we have developed a novel nanoimmunotherapy that combines locally administered Prussian blue nanoparticle-based photothermal therapy(PBNP-PTT)with systemically administered agonistic anti-CD137 monoclonal antibody therapy(aCD137).PBNP-PTT was administered at various thermal doses to melanoma cells in vitro,and was combined with aCD137 in vivo to test treatment effects on melanoma tumor progression,animal survival,immunological protection against tumor rechallenge,and *** administered at a melanoma-specific thermal dose,PBNP-PTT elicits immunogenic cell death(ICD)in melanoma cells and upregulates markers associated with antigen presentation and immune cell co-stimulation in ***,PBNP-PTT eliminates primary melanoma tumors in vivo,yielding long-term tumor-free ***,the antitumor immune effects generated by PBNP-PTT cannot eliminate secondary tumors,despite significantly slowing their *** addition of aCD137 enables significant abscopal efficacy and improvement of survival,functioning through activated dendritic cells and tumor-infiltrating CD8^(+)T cells,and generates CD4^(+)and CD8^(+)T cell memory that manifests in the rejection of tumor rechallenge,with no long-term *** study describes for the first time a novel and effective nanoimmunotherapy combination of PBNP-PTT with aCD137 mAb therapy for melanoma.

关键词： photothermal therapy Prussian blue nanoparticles anti-CD137 melanoma nanoimmunotherapy

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Stabilization of Alliinase from Garlic by Osmolytes and the Mannose-Specific Lectin ASAI

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Journal of Pharmacy and Pharmacology 2018年第5期6卷 437-447页

作者： Irina Shin David Mirelman Lev Weiner Enrique Villar Valery L. Shnyrov Aharon Rabinkov Department of Life Sciences core facilities Weizmann Institute of Science Rehovot 76100 Israel Department of Biomolecular Science Weizmann Institute of Science Rehovot 76100 Israel Department of Chemical Research Support Weizmann Institute of Science Rehovot 76100 Israel Department of biochemistry and molecular Biology Universidad de Salamanca Salamanca 37007 Spain

Objectives： Alliinase is a pyridoxal-5＇-phosphate （PLP）-dependent enzyme responsible for the production of diallyl thiosulfinate （allicin）, the biologically active component of garlic, from alliin. The use of allicin for treatment of various diseases has been proposed but it is very unstable in the blood stream. This difficulty can be overcome by administration of alliin, together with a conjugate of alliinase directed towards the target cells. This, in turn requires a stable and active form of the enzyme. In this study we evaluate the stability of alliinase itself, in the presence mad absence of osmolytes, as well as that of its catalytically active complex with a mannose-specific lectin, ASAI （Allium sativum agglutinin I）, also presents in garlic. Methods： Alliinase, mad ASAI were both purified from garlic cloves. Thermal stability of alliinase itself, mad of its complexes with PLP and ASAI, in the presence mad absence of osmolytes, was analyzed by monitoring enzymic activity, and using DSC （differential scanning calorimetry）. Key findings： PLP exerts only a minor influence on alliinase structure and stablity. But both osmolytes and ASAI stabilize the enzyme considerably. Conclusions： The principle finding is that ASAI greatly stabilizes alliinase. Thus, the lectin-enzyme complex, which can be lyophilized and stored until used, provides an effective formulation of alliinase for generation of allicin from alliin in vivo.

关键词： Alliinase osmolyte mannose-specific lectin ASAI pyridoxal 5'-phosphate.

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The GYF domain protein CD2BP2 is critical for embryogenesis and podocyte function

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分子细胞生物学报:英文版 2015年第5期7卷 402-414页

作者： Gesa I.Albert Christoph Schell Karin M.Kirschner Sebastian Schafer Ronald Naumann Alexandra Muller Oliver Kretz Benno Kuropka Mathias Girbig Norbert Hubner Eberhard Krause Holger Scholz Tobias B.Huber Klaus-Peter Knobeloch Christian Freund Institute for Chemistry and biochemistry Freie Universitat Berlin14195 BerlinGermany Leibniz-Institut fuer Molekulare Pharmakologie 13125 BerlinGermany Renal Division University Hospital Freiburg79106 FreiburgGermany Speman Graduate School for medicine and Biology University of Freiburg79106 FreiburgGermany Faculty of Biology University Freiburg79106 FreiburgGermany Institute fuer Vegetative Physiologie Charité-Universitatsmedizin Berlin10117 BerlinGermany Experimental Genetics and Cardiovascular Diseases MDC13125 BerlinGermany Transgenic core facility Max-Planck-Institute of Molecular Cell Biology and GeneticsDresdenGermany Institute of Neuropathology University of Freiburg79106 FreiburgGermany Institute of Cell Biology&Anatomy University of Freiburg79106 FreiburgGermany BIOSS Centre for Biological Signaling Studies Albert-Ludwigs-University79106 FreiburgGermany

Scaffolding proteins play pivotal roles in the assembly of macromolecular machines such as the *** adaptor protein CD2BP2,originally identified as a binding partner of the adhesion molecule CD2,is a pre-spliceosomal assembly factor that utilizes its glycine-tyrosine-phenylalanine(GYF)domain to co-localize with spliceosomal *** far,its function in vertebrates is *** conditional gene targeting in mice,we show that CD2BP2 is crucial for embryogenesis,leading to growth retardation,defects in vascularization,and premature death at embryonic day 10.5 when *** of the protein in bone marrow-derived macrophages indicates that CD2BP2 is involved in the alternative splicing of mRNA transcripts from diverse *** the molecular level,we identified the phosphatase PP1 to be recruited to the spliceosome via the N-terminus of *** the strong expression of CD2BP2 in podocytes of the kidney,we use selective depletion of CD2BP2,in combination with next-generation sequencing,to monitor changes in exon usage of genes critical for podocyte functions,including VEGF and actin ***2BP2-depleted podocytes display foot process effacement,and cause proteinuria and ultimately lethal kidney failure in ***,our study defines CD2BP2 as a non-redundant splicing factor essential for embryonic development and podocyte integrity.

关键词： GYF domain CD2BP2 alternative splicing PP1 podocytes VEGF

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Monoclonal Antibodies Reveal Novel Localization of SPAG11E in Spermatozoa and the Antifertility Potential of SPAG11E Motifs

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Reproductive and Developmental medicine 2018年第4期2卷 191-200,I0001-I0005页

作者： Zhi-Kai Wang Yi-Ting Yang Xin-Yu Chen Shuang-Gang Hu Ping Zhu Wan-Xiang Xu Li Ma He-Guo Yu Hua Diao Yong-Lian Zhang Clinical Laboratory(Hospital of the SIPPR)and NHC Key Laboratory of Reproduction Regulation Shanghai Institute of Planned Parenthood ResearchShanghai 200032China core facility of Basic Medical Sciences School of MedicineShanghai Jiao Tong UniversityShanghai 200025China Shanghai Key Laboratory of molecular Andrology State Key Laboratory of Molecular BiologyInstitute of Biochemistry and Cell BiologyChinese Academy of SciencesShanghai 200031China

Objective: SPAG11E is the first β-defensin that has been reported to activate Ca;uptake and sperm motility. However, the exact subcellular localization and interaction of SPAG11E with sperm remain controversial because of the lack of qualified antibody tools. SPAG11E is also a potential male antifertility target because SPAG11E fragment conjugated with a carrier protein exhibits male contraceptive vaccine ***, the fine B-cell epitope motifs of SPAG11E have not been analyzed, which hampered further exploration of the potential ***: Polyclonal and monoclonal antibodies（mcAbs） of mature SPAG11E were raised and qualified with Western blotting. Subcellular localization of SPAG11E was revealed by Western blotting, immunohistochemistry staining, and electron microscopy. B-cell epitopes of rat SPAG11E were mapped by Western blotting using polyclonal and mcAbs. Based on the conservation of the identified epitope motifs between rat and mouse SPAG11E, antifertility potential of the epitope motifs was evaluated by the offspring of the males compromised with specific ***: SPAG11E antibodies of high quality were obtained and all B-cell epitope motifs of rat SPAG 11 E were mapped, in which conserved epitope motifs of SPAG11E in various species were discovered. The epitope motifs recognized by mcAbs were identified respectively. With mcAbs, rat SPAG11E was proved to be expressed in the caput region of epididymis. A novel finding was that SPAG 11 E was located in the flagella and nuclei of sperm as revealed by immunoelectron microscopy. In addition, the males treated with mcAbs（3#-1 and 10#B4） showed apparently fewer ***: SPAG11E revealed a β-defensin with novel localization in sperm flagellum and nucleus with qualified antibodies. All B-cell epitope motifs of rat SPAG11E were determined, and the antifertility potential was proved by corresponding mcAbs.

关键词： Antibody Contraception Epitope SPAG11E Sperm

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Interaction of the α2A domain of integrin with small collagen fragments

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Protein & Cell 2010年第4期1卷 393-405页

作者： Hans-Christian Siebert Monika Burg-Roderfeld Thomas Eckert Sabine Stötzel Ulrike Kirch Tammo Diercks Martin J.Humphries Martin Frank Rainer Wechselberger Emad Tajkhorshid Steffen Oesser Institut für Biochemie und Endokrinologie Veterinärmedizinische FakultätJustus-Liebig-Universität GießenFrankfurter Str.10035392 GießenGermany CiC bioGUNE Parque Tecnológico de BizkaiaEdificio 80048160 DerioSpain Utrecht facility for High-resolution NMR Bijvoetcenter for Biomolecular Research Utrecht UniversityPadualaan 83584CH UtrechtThe Netherlands Wellcome Trust Centre for Cell-Matrix Research School of Biological SciencesUniversity of Manchester2.205 Stopford BuildingOxford RoadManchester M139PTUK molecular Structure Analysis core facility Deutsches KrebsforschungszentrumIm Neuenheimer Feld 28069120 HeidelbergGermany Department of biochemistry Beckman Instituteand Center for Biophysics and Computational BiologyUniversity of Illinois at Urbana-ChampaignUrbanaILUSA Collagen Research Institute Schauenburgerstr.116D-24118 KielGermany

We here present a detailed study of the ligand-receptor interactions between single and triple-helical strands of collagen and the α2A domain of integrin(α2A),providing valuable new insights into the mechanisms and dynamics of collagen-integrin binding at a sub-molecular *** occurrence of single and triple-helical strands of the collagen fragments was scrutinized with atom force microscopy(AFM)*** interactions of the triple-stranded fragments comparable to those of collagen can only be detected for the 42mer triple-helical collagen-like peptide under study(which contains 42 amino acid residues per strand)by solid phase assays as well as by surface plasmon resonance(SPR)***,changes in NMR signals during titration and characteristic saturation transfer difference(STD)NMR signals are also detectable whenα2A is added to a solution of the 21mer single-stranded collagen *** dynamics(MD)simulations employing different sets of force field parameters were applied to study the interaction between triple-helical or single-stranded collagen fragments withα*** is remarkable that even single-stranded collagen fragments can form various complexes withα2A showing significant differences in the complex stability with identical *** results of MD simulations are in agreement with the signal alterations in our NMR experiments,which are indicative of the formation of weak complexes between single-stranded collagen andα2A in *** results provide useful information concerning possible interactions ofα2A with small collagen fragments that are of relevance to the design of novel therapeutic A-domain inhibitors.

关键词： integrin-collagen interaction NMR SPR AFM molecular modeling

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TCTE1 is a conserved component of the dynein regulatory complex and is required for motility and metabolism in mouse spermatozoa

TCTE1 is a conserved component of the dynein regulatory comp...

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中国生理学会生殖科学专业委员会—中国动物学会生殖生物学分会第二次联合学术年会暨“生殖科学专业委员会第二次学术交流会”和“生殖生物学分会第十六次学术年会”

作者： Julio M.Castaneda Rong Hua Haruhiko Miyata Asami Oji Yueshuai Guo Yiwei Cheng Tao Zhou Xuejiang Guo Yiqiang Cui Bin Shen Zibin Wang Zhibin Hu Zuomin Zhou Jiahao Sha Renata Prunskaite-Hyyrylainen Zhifeng Yu Ramiro Ramirez-Solis Masahito lkawa Martin M.Matzuk Mingxi Liu Department of Pathology and Immunology Baylor College of Medicine Research Institute for Microbial Diseases Osaka University State Key Laboratory of Reproductive medicine Nanjing Medical University Department of Histology and Embryology Nanjing Medical University Graduate School of Pharmaceutical Sciences Osaka University Animal core facility of Nanjing Medical University Center for Reproductive medicine Baylor College of Medicine Faculty of biochemistry and molecular medicine University of Oulu Center for Drug Discovery Baylor College of Medicine Wellcome Trust Sanger Institute The Institute of Medical Science The University of Tokyo Department of molecular and Cellular Biology Baylor College of Medicine Department of molecular and Human Genetics Baylor College of Medicine Department of Pharmacology Baylor College of Medicine

Flagella and cilia are critical cellular organelles that provide a means for cells to sense and progress through their *** central component of flagella and cilia is the axoneme,which comprises the"9+2"microtubule arrangement,dynein arms,radial spokes,and the nexin-dynein regulatory complex(N-DRC).Failure to properly assemble components of the axoneme leads to defective flagella and in humans leads to

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LIN28 coordinately promotes nucleolar/ribosomal functions and represses the 2C-like transcriptional program in pluripotent stem cells

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Protein & Cell 2022年第7期13卷 490-512页

作者： Zhen Sun Hua Yu Jing Zhao Tianyu Tan Hongru Pan Yuqing Zhu Lang Chen Cheng Zhang Li Zhang Anhua Lei Yuyan Xu Xianju Bi Xin Huang Bo Gao Longfei Wang Cristina Correia Ming Chen Qiming Sun Yu Feng Li Shen Hao Wu Jianlong Wang Xiaohua Shen George Q.Daley Hu Li Jin Zhang Center for Stem Cell and Regenerative medicine Department of Basic Medical Sciences and the First Affiliated HospitalZhejiang University School of MedicineHangzhou 310058China Department of molecular Pharmacology&Experimental Therapeutics Center for Individualized MedicineMayo ClinicRochesterMNUSA Tsinghua-Peking Center for Life Sciences School of MedicineTsinghua UniversityBeijing 100085China The Black Family Stem Cell Institute and Department of Cell Developmental and Regenerative BiologyIcahn School of Medicine at Mount SinaiNew YorkNY 10029USA Department of Biophysics and Department of Pathology of Sir Run Run Shaw Hospital Zhejiang University School of MedicineHangzhou 31005&China Department of Biological Chemistry and molecular Pharmacology Harvard Medical Schooland Program in Cellular and Molecular MedicineBoston Children's HospitalBostonMAUSA Stem Cell Transpla ntation Program Division of Pediatric Hematology On cologyBost on Children's HospitalDepartme nt of Biological Chemistry and Molecular PharmacologyHarvard Medical SchoolBostonMAUSA College of Life Sciences Zhejiang UniversityHangzhou 310058China Department of biochemistry Zhejiang University School of MedicineHangzhou 310058China Institute of Life Science Zhejiang UniversityHangzhou 310058China Institute of Hematology Zhejiang UniversityHangzhou 310058China Zhejiang Laboratory for Systems and Precision medicine Zhejiang University Medical CenterHangzhou 310058China

LIN28 is an RNA binding protein with important roles in early embryo development,stem cell differentiation/re-programming,tumorigenesis and *** studies have focused mainly on its role in the cytosol where it interacts with Let-7 microRNA precursors or mRNAs,and few have addressed LIN28's role within the ***,we show that LIN28 displays dynamic temporal and spatial expression during murine embryo *** LIN28 expression drops upon exit from the 2-cell stage,and zygotic LIN28 protein is induced at the forming nucleolus during 4-cell to blas-tocyst stage development,to become dominantly expressed in the cytosol after *** cultured pluripotent stem cells(PSCs),loss of LIN28 led to nucleolar stress and activation of a 2-cell/4-cell-like transcriptional program characterized by the expression of endogenous retrovirus ***,LIN28 binds to small nucleolar RNAs and rRNA to maintain nucleolar integrity,and its loss leads to nucleolar phase separation defects,ribosomal stress and activation of P53 which in turn binds to and activates 2C transcription factor ***28 also resides in a complex containing the nucleolar factor Nucleolin(NCL)and the transcrip-tional repressor TRIM28,and LIN28 loss leads to reduced occupancy of the NCL/TRIM28 complex on the Dux and rDNA loci,and thus de-repressed Dux and reduced rRNA ***28 knockout cells with nucleolar stress are more likely to assume a slowly cycling,translationally inert and anabolically inactive state,which is a part of previously unappreciated 2C-like transcriptional *** findings elucidate novel roles for nucleolar LIN28 in PSCs,and a new mechanism linking 2C program and nucleolar functions in PSCs and early embryo development.

关键词： LIN28 2-cell-like program nucleolar integrity NCL/TRIM28 complex

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