DOCSLIB.ORG

Ninth UK Neuromuscular Translational Research Conference 22Nd – 23Rd March 2016

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Ninth UK Neuromuscular Translational Research Conference 22Nd – 23Rd March 2016 Ninth UK Neuromuscular Translational Research Conference 22nd – 23rd March 2016 Medical Sciences Teaching Centre Oxford Oxford, OX1 3PL Table of Contents Welcome from the MRC Centre for Neuromuscular Diseases and from Muscular Dystrophy UK .......................................................................................... 2 About the MRC Centre for Neuromuscular Diseases and Muscular Dystrophy UK ............ 5 Patient organisations ............................................................................................. 8 Programme .......................................................................................................... 9 Flash poster list ................................................................................................... 14 Poster list ........................................................................................................... 15 Invited speaker abstracts ...................................................................................... 24 Poster and platform presentation abstracts .............................................................. 30 Clinical trials in the MRC Centre for Neuromuscular Diseases ................................... 111 Delegate list ...................................................................................................... 131 MRC Centre for Neuromuscular Diseases staff list ................................................... 140 Conference Planning Group ................................................................................. 143 MDUK supported Principal Investigators ................................................................ 144 Educational opportunities .................................................................................... 145 Sponsors ........................................................................................................... 147 1 Welcome to the UK Neuromuscular Translational Research Conference, Oxford 2016 Dear Colleagues, We are delighted to welcome you to Oxford for the ninth annual UK Neuromuscular Translational Research Conference. We are very pleased that this conference continues to be jointly hosted by the MRC Centre for Neuromuscular Diseases and Muscular Dystrophy UK. In addition, this year we have worked closely with colleagues in the London-Newcastle MRC Centre steering committee, the Wellcome Trust Centre for Mitochondrial Research in Newcastle and the MRC Functional Genomics unit in Oxford to develop the scientific translational research programme. Major translational themes this year include a wide ranging session on genomic therapies, an in-depth review of next generation biomarkers and a session tackling the difficult issue of defining and how to deal with big data. We are delighted to welcome Professor David Beeson from the University of Oxford to deliver the third John Newsom-Davis lecture. Professor Beeson’s renowned work on inherited neuromuscular junction disorders makes him the ideal person to deliver this lecture. After our inaugural and very successful patient day at our conference in Newcastle last year, we are running another patient day this year the day before the conference itself. The mission of the MRC Centre is to translate science into new experimental medicine trials and find treatments for children and adults with serious muscle wasting diseases. Collaborative working and interdisciplinarity represent a fundamental platform for successful translation. The MRC Centre works to bring together clinicians, scientists, patient organisations and patients in order to advance the UK translational research effort. This is a particularly exciting time in the field as scientific discoveries are revealing an increasing number of tractable therapeutic targets. The MRC Centre continues to work closely with all its partners to support the development of a clinical trials culture and aims to embed an experimental clinical trials network in neuromuscular clinical practice. We will continue to work hard to form effective research and clinical links with as many other UK neuromuscular groups as possible. In the renewed Centre (renewed 2013-2018) we have developed strong experimental medicine links with, and received important coordinated support from, three NHS NIHR Biomedical Research Centres based at University College London Hospitals, Great Ormond Street and at Newcastle University Hospitals. The MRC Centre is also pleased to contribute to the NIHR National Rare Diseases Translational Research Collaboration. The MRC Centre was established in 2008 as a joint partnership between the UCL Institute of Neurology, Queen Square, the UCL Institute of Child Health and the Newcastle University and was renewed for a further five years in 2013. The Centre continues to be closely linked to its partner NHS organisations, University College London Hospitals NHS Foundation Trust, Great Ormond Street Hospital for Children NHS Foundation Trust and Newcastle upon Tyne Hospitals NHS Foundation Trust. The Centre has also developed strong links with groups in Oxford and Cambridge which we continue to build on in this current phase of the Centre. Over the next two days this conference aims to showcase a wide range of high quality scientific neuromuscular research from many UK groups, international colleagues and industry partners. 2 We have received 125 high quality abstracts, and there will be platform presentations and dedicated poster sessions each day as well as guided poster discussions. This year will be the second time we add a poster flash session where abstracts are selected to be presented in 5 minute slots using only three slides. We hope this will prove a popular session. There will be five £500 poster prizes for young investigators. Accepted abstracts will be published in a special supplement of the journal Neuromuscular Disorders with Professor Mary M. Reilly as the special guest editor. As the Centre Director, I would very much like to thank Professor Mary M. Reilly who has led the joint MRC-Muscular Dystrophy UK meeting scientific planning team: Professors Kate Bushby, Doug Turnbull, Francesco Muntoni, Professor Dame Kay E Davies, Professor Kevin Talbot and Dr Marita Pohlschmidt. I also sincerely thank Christine Oldfield and Dr Marita Pohlschmidt for their very hard work in organising this meeting. I am very grateful for the interest, sponsorship and support of industry colleagues who are working with us to develop new therapies for patients with neuromuscular diseases. Once again this annual meeting has been oversubscribed. We are very encouraged that there continues to be such strong interest in neuromuscular translational research from throughout the UK and beyond. We sincerely hope that you have a stimulating, productive and entertaining two days in Oxford. Professor Michael G. Hanna Professor Mary M. Reilly Director Co-Director MRC Centre for Neuromuscular Diseases MRC Centre for Neuromuscular Diseases UCL Institute of Neurology UCL Institute of Neurology Professor Francesco Muntoni Professor Katie Bushby Co-Director, ICH/GOS Co-Director, Newcastle MRC Centre for Neuromuscular Diseases MRC Centre for Neuromuscular Diseases UCL Institute of Neurology Newcastle University Professor Doug Turnbull Professor Dame Kay E. Davies Co-Director Director, MRC Functional Genomics Unit MRC Centre for Neuromuscular Diseases University of Oxford Newcastle University Dr Marita Pohlschmidt Professor Kevin Talbot Director of Research, Head of the Division of Clinical Neurology Muscular Dystrophy UK University of Oxford 3 Welcome to the Conference Robert Meadowcroft, Chief Executive of Muscular Dystrophy UK I am delighted to welcome you to Oxford and the 2016 UK Neuromuscular Translational Research Conference. We have expert speakers, interesting research posters and a great opportunity to develop and share ideas and I trust you enjoy your time here. Muscular Dystrophy UK has supported this important conference since its inception and we welcome and encourage the opportunity it presents to bring scientists and clinical researchers together each year. As a funder of research, we have invested more than £55million in backing cutting-edge research into neuromuscular conditions. We also make a major investment each year in improving specialist NHS care in the UK as well providing information, advice and support for families. We are planning to increase our support in the coming years of high-quality science and translational research here in the UK and through international partnerships to address a number of the rarer conditions such as nemaline myopathy and the Collagen VI disorders. We share the concern of many families and supporters that every day counts in the race to develop effective treatments particularly for the more aggressive conditions that affect children. For many adults with a muscle-wasting condition, there is a similar focus on research and also an understanding that expert clinical care, retaining independence and enjoying the best possible quality of life are all important goals. We are encouraged by the significant progress being made towards ensuring effective treatments although there are many questions that remain unanswered. We need to develop and refine the use of MRI and also develop additional outcome measures. The heterogeneity of many conditions underlines the need for better characterisation and an improved understanding of the natural history. Nevertheless, we are very encouraged by the increase, year on year, in the number of clinical trials
Load more

Recommended publications
  • Necroptosis in Intestinal Inflammation and Cancer
    biomolecules Review Necroptosis in Intestinal Inflammation and Cancer: New Concepts and Therapeutic Perspectives Anna Negroni 1,* , Eleonora Colantoni 2, Salvatore Cucchiara 2 and Laura Stronati 3 1 Division of Health Protection Technologies, ENEA, 00123 Rome, Italy 2 Maternal Infantile and Urological Sciences Department, Sapienza, University of Rome, 00161 Rome, Italy; [email protected] (E.C.); [email protected] (S.C.) 3 Department of Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-06-3048-3623 Received: 3 September 2020; Accepted: 8 October 2020; Published: 10 October 2020 Abstract: Necroptosis is a caspases-independent programmed cell death displaying intermediate features between necrosis and apoptosis. Albeit some physiological roles during embryonic development such tissue homeostasis and innate immune response are documented, necroptosis is mainly considered a pro-inflammatory cell death. Key actors of necroptosis are the receptor-interacting-protein-kinases, RIPK1 and RIPK3, and their target, the mixed-lineage-kinase-domain-like protein, MLKL. The intestinal epithelium has one of the highest rates of cellular turnover in a process that is tightly regulated. Altered necroptosis at the intestinal epithelium leads to uncontrolled microbial translocation and deleterious inflammation. Indeed, necroptosis plays a role in many disease conditions and inhibiting necroptosis is currently considered a promising therapeutic strategy. In this review, we focus on the molecular mechanisms of necroptosis as well as its involvement in human diseases. We also discuss the present developing therapies that target necroptosis machinery. Keywords: programmed cell death; inflammation; cancer; intestinal diseases; inhibitors 1. Introduction Cell death is crucial during the development and maintenance of tissue homeostasis in multicellular organisms.
    [Show full text]
  • Global Survey of Cell Death Mechanisms Reveals Metabolic Regulation of Ferroptosis
    Global Survey of Cell Death Mechanisms Reveals Metabolic Regulation of GPX4-Dependent Ferroptosis Kenichi Shimada Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2015 ©2015 Kenichi Shimada All rights reserved ABSTRACT Global Survey of Cell Death Mechanisms Reveals Metabolic Regulation of Ferroptosis Kenichi Shimada Cells die not merely as a consequence of catastrophic failure of homeostasis but when programmed cell death is activated. The existence of non-apoptotic modes of regulated cell death is increasingly appreciated. However, the full extent and diversity of these alternative cell death mechanisms remains uncharted. In this thesis, we developed a systematic framework to discover and characterize lethal compounds that induce distinct cell death phenotypes. In the first part, we investigated the landscape of pharmacologically-accessible non-apoptotic cell death mechanisms. This effort resulted in the discovery of a novel ferroptosis inducer, FIN56. The rest of my work focused on characterizing the mechanism of action of FIN56. Technologies used here should be generally applicable for the systematic study of various cell death mechanisms. First, to globally survey pharmacologically-accessible cell death mechanisms, we used 3,169 uncharacterized lethal compounds as cell death probes. We found that 451 compounds (14%) were lethal without activating caspase activity. 56 most potent and structurally diverse compounds were more closely studied using the 'modulatory profiling' approach, which involves examining changes in potency of lethal compounds by co-treatment with chemical modulators. We discovered that caspase-independent lethals induced three types of regulated non-apoptotic cell death: metal ion-dependent cell death, necrostatin-1-dependent cell death, and ferroptosis, a regulated form of iron-dependent oxidative cell death.
    [Show full text]
  • Regulation of Caspase-8 Activity at the Crossroads of Pro-Inflammation
    International Journal of Molecular Sciences Review Regulation of Caspase-8 Activity at the Crossroads of Pro-Inflammation and Anti-Inflammation Jun-Hyuk Han 1, Jooho Park 1,2, Tae-Bong Kang 1,3,* and Kwang-Ho Lee 1,3 1 Department of Applied Life Sciences, Graduate School, BK21 Program, Konkuk University, Chungju 27478, Korea; [email protected] (J.-H.H.); [email protected] (J.P.); [email protected] (K.-H.L.) 2 Department of Biomedical Chemistry, College of Biomedical & Health Science, Konkuk University, Chungju 27487, Korea 3 Department of Biotechnology, College of Biomedical & Health Science, Konkuk University, Chungju 27487, Korea * Correspondence: [email protected]; Tel.: +82-43-840-3904 Abstract: Caspase-8 has been classified as an apoptotic caspase, and its initial definition was an initiator of extrinsic cell death. During the past decade, the concept of caspase-8 functioning has been changed by findings of its additional roles in diverse biological processes. Although caspase-8 was not originally thought to be involved in the inflammation process, many recent works have determined that caspase-8 plays an important role in the regulatory functions of inflammatory processes. In this review, we describe the recent advances in knowledge regarding the manner in which caspase-8 modulates the inflammatory responses concerning inflammasome activation, cell death, and cytokine induction. Keywords: caspase-8; inflammasome; inflammation; necroptosis; pyroptosis; apoptosis Citation: Han, J.-H.; Park, J.; Kang, T.-B.; Lee, K.-H. Regulation of Caspase-8 Activity at the Crossroads 1. Introduction of Pro-Inflammation and Anti-Inflammation. Int. J. Mol. Sci. Mammalian caspases have classically been divided into inflammatory and apoptotic 2021, 22, 3318.
    [Show full text]
  • Molecular Insights Into the Mechanism of Necroptosis: the Necrosome As a Potential Therapeutic Target
    cells Review Molecular Insights into the Mechanism of Necroptosis: The Necrosome as a Potential Therapeutic Target 1, 1, 1,2 1, Jing Chen y, Renate Kos y , Johan Garssen and Frank Redegeld * 1 Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, 3508 TB Utrecht, The Netherlands; [email protected] (J.C.); [email protected] (R.K.); [email protected] (J.G.) 2 Danone Nutricia Research, Uppsalaan 12, 3584 CT Utrecht, The Netherlands * Correspondence: [email protected]; Tel.: +31-6-20252139 Jing Chen and Renate Kos share first authorship. y Received: 30 October 2019; Accepted: 19 November 2019; Published: 21 November 2019 Abstract: Necroptosis, or regulated necrosis, is an important type of programmed cell death in addition to apoptosis. Necroptosis induction leads to cell membrane disruption, inflammation and vascularization. It plays important roles in various pathological processes, including neurodegeneration, inflammatory diseases, multiple cancers, and kidney injury. The molecular regulation of necroptotic pathway has been intensively studied in recent years. Necroptosis can be triggered by multiple stimuli and this pathway is regulated through activation of receptor-interacting protein kinase 1 (RIPK1), RIPK3 and pseudokinase mixed lineage kinase domain-like (MLKL). A better understanding of the mechanism of regulation of necroptosis will further aid to the development of novel drugs for necroptosis-associated human diseases. In this review, we focus on new insights in the regulatory machinery of necroptosis. We further discuss the role of necroptosis in different pathologies, its potential as a therapeutic target and the current status of clinical development of drugs interfering in the necroptotic pathway.
    [Show full text]
  • Caspase-8 Promotes C-Rel–Dependent Inflammatory Cytokine Expression and Resistance Against Toxoplasma Gondii
    Caspase-8 promotes c-Rel–dependent inflammatory cytokine expression and resistance against Toxoplasma gondii Alexandra A. DeLaneya,b, Corbett T. Berrya,c,1, David A. Christiana,1, Andrew Hartd, Elisabet Bjanesa,b, Meghan A. Wynosky-Dolfia, Xinyuan Lie, Bart Tummersf, Irina A. Udalovag, Youhai H. Chene,h, Uri Hershbergc, Bruce D. Freedmana,b,d,h, Christopher A. Huntera,b,d,h, and Igor E. Brodskya,b,d,h,2 aDepartment of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA 19104; bCell and Molecular Biology Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104; cSchool of Biomedical Engineering, Drexel University College of Medicine, Philadelphia, PA 19104; dImmunology Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104; eDepartment of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104; fDepartment of Immunology, St. Jude Children’s Research Hospital, Memphis, TN 38105; gKennedy Institute of Rheumatology, Medical Sciences Division, University of Oxford, OX3 9DU Oxford, United Kingdom; and hInstitute for Immunology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104 Edited by Ruslan Medzhitov, Yale University School of Medicine, New Haven, CT, and approved May 2, 2019 (received for review December 4, 2018) Caspase-8 is a key integrator of cell survival and cell death decisions cell-extrinsic apoptosis in response to various death-inducing stimuli, during infection and inflammation. Following engagement of tumor such as Fas (15) and tumor necrosis factor (TNF) (16), but also necrosis factor superfamily receptors or certain Toll-like receptors has a prosurvival function in concert with its enzymatically in- (TLRs), caspase-8 initiates cell-extrinsic apoptosis while inhibiting active homolog cFLIP.
    [Show full text]
  • Conformational Interconversion of MLKL and Disengagement from RIPK3 Precede Cell Death by Necroptosis
    ARTICLE https://doi.org/10.1038/s41467-021-22400-z OPEN Conformational interconversion of MLKL and disengagement from RIPK3 precede cell death by necroptosis Sarah E. Garnish1,2,8, Yanxiang Meng 1,2,8, Akiko Koide3,4,8, Jarrod J. Sandow 1,2, Eric Denbaum3, Annette V. Jacobsen 1,2, Wayland Yeung5, Andre L. Samson 1,2, Christopher R. Horne 1,2, Cheree Fitzgibbon1, Samuel N. Young1, Phoebe P. C. Smith 1, Andrew I. Webb 1,2, Emma J. Petrie1,2, ✉ Joanne M. Hildebrand 1,2, Natarajan Kannan5,6, Peter E. Czabotar 1,2, Shohei Koide 3,7,9 & ✉ James M. Murphy 1,2,9 1234567890():,; Phosphorylation of the MLKL pseudokinase by the RIPK3 kinase leads to MLKL oligomer- ization, translocation to, and permeabilization of, the plasma membrane to induce necroptotic cell death. The precise choreography of MLKL activation remains incompletely understood. Here, we report Monobodies, synthetic binding proteins, that bind the pseudokinase domain of MLKL within human cells and their crystal structures in complex with the human MLKL pseudokinase domain. While Monobody-32 constitutively binds the MLKL hinge region, Monobody-27 binds MLKL via an epitope that overlaps the RIPK3 binding site and is only exposed after phosphorylated MLKL disengages from RIPK3 following necroptotic stimula- tion. The crystal structures identified two distinct conformations of the MLKL pseudokinase domain, supporting the idea that a conformational transition accompanies MLKL disen- gagement from RIPK3. These studies provide further evidence that MLKL undergoes a large conformational change upon activation, and identify MLKL disengagement from RIPK3 as a key regulatory step in the necroptosis pathway.
    [Show full text]
  • Network-Assisted Analysis of Primary Sjögren's Syndrome GWAS Data In
    Network-assisted analysis of primary Sjögren’s syndrome GWAS data in Han Chinese Kechi Fang, Kunlin Zhang, Jing Wang* Address: Key Laboratory of Mental Health, Institute of Psychology, Chinese Academy of Sciences, Beijing 100101, China. Email: Kechi Fang – [email protected]; Kunlin Zhang – [email protected]; Jing Wang – [email protected] *Corresponding author 1 Supplementary materials Page 3 – Page 5: Supplementary Figure S1. The direct network formed by the module genes from DAPPLE. Page 6: Supplementary Figure S2. Transcript expression heatmap. Page 7: Supplementary Figure S3. Transcript enrichment heatmap. Page 8: Supplementary Figure S4. Workflow of network-assisted analysis of pSS GWAS data to identify candidate genes. Page 9 – Page 734: Supplementary Table S1. A full list of PPI pairs involved in the node-weighted pSS interactome. Page 735 – Page 737: Supplementary Table S2. Detailed information about module genes and sigMHC-genes. Page 738: Supplementary Table S3. GO terms enriched by module genes. 2 NFKBIE CFLAR NFKB1 STAT4 JUN HSF1 CCDC90B SUMO2 STAT1 PAFAH1B3 NMI GTF2I 2e−04 CDKN2C LAMA4 8e−04 HDAC1 EED 0.002 WWOX PSMD7 0.008 TP53 PSMA1 HR 0.02 RPA1 0.08 UBC ARID3A PTTG1 0.2 TSC22D4 ERH NIF3L1 0.4 MAD2L1 DMRTB1 1 ERBB4 PRMT2 FXR2 MBL2 CBS UHRF2 PCNP VTA1 3 DNMT3B DNMT1 RBBP4 DNMT3A RFC3 DDB1 THRA CBX5 EED NR2F2 RAD9A HUS1 RFC4 DDB2 HDAC2 HCFC1 CDC45L PPP1CA MLLSMARCA2 PGR SP3 EZH2 CSNK2B HIST1H4C HIST1H4F HNRNPUL1 HR HIST4H4 TAF1C HIST1H4A ENSG00000206300 APEX1 TFDP1 RHOA ENSG00000206406 RPF2 E2F4 HIST1H4IHIST1H4B HIST1H4D
    [Show full text]
  • RIPK3 / RIP3 Antibody (Aa480-530) Rabbit Polyclonal Antibody Catalog # ALS11903
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 RIPK3 / RIP3 Antibody (aa480-530) Rabbit Polyclonal Antibody Catalog # ALS11903 Specification RIPK3 / RIP3 Antibody (aa480-530) - Product Information Application IHC Primary Accession Q9Y572 Reactivity Human Host Rabbit Clonality Polyclonal Calculated MW 57kDa KDa RIPK3 / RIP3 Antibody (aa480-530) - Additional Information Gene ID 11035 Anti-RIPK3 / RIP3 antibody IHC of human Other Names pancreas. Receptor-interacting serine/threonine-protein kinase 3, 2.7.11.1, RIP-like protein kinase 3, RIPK3 / RIP3 Antibody (aa480-530) - Receptor-interacting protein 3, RIP-3, RIPK3, Background RIP3 Essential for necroptosis, a programmed cell Target/Specificity death process in response to death-inducing A portion of amino acids 480-518 of human TNF-alpha family members. Upon induction of RIP3 necrosis, RIPK3 interacts with, and phosphorylates RIPK1 and MLKL to form a Reconstitution & Storage necrosis-inducing complex. RIPK3 binds to and Short term 4°C, long term aliquot and store enhances the activity of three metabolic at -20°C, avoid freeze thaw cycles. enzymes: GLUL, GLUD1, and PYGL. These metabolic enzymes may eventually stimulate Precautions the tricarboxylic acid cycle and oxidative RIPK3 / RIP3 Antibody (aa480-530) is for research use only and not for use in phosphorylation, which could result in diagnostic or therapeutic procedures. enhanced ROS production. RIPK3 / RIP3 Antibody (aa480-530) - References RIPK3 / RIP3 Antibody (aa480-530) - Protein Information Yu P.W.,et al.Curr. Biol. 9:539-542(1999). Sun X.,et al.J. Biol. Chem. Name RIPK3 (HGNC:10021) 274:16871-16875(1999).
    [Show full text]
  • Mouse Ripk3 Conditional Knockout Project (CRISPR/Cas9)
    https://www.alphaknockout.com Mouse Ripk3 Conditional Knockout Project (CRISPR/Cas9) Objective: To create a Ripk3 conditional knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering. Strategy summary: The Ripk3 gene (NCBI Reference Sequence: NM_001164108 ; Ensembl: ENSMUSG00000022221 ) is located on Mouse chromosome 14. 10 exons are identified, with the ATG start codon in exon 3 and the TAG stop codon in exon 10 (Transcript: ENSMUST00000168716). Exon 3~9 will be selected as conditional knockout region (cKO region). Deletion of this region should result in the loss of function of the Mouse Ripk3 gene. To engineer the targeting vector, homologous arms and cKO region will be generated by PCR using BAC clone RP24-66O7 as template. Cas9, gRNA and targeting vector will be co-injected into fertilized eggs for cKO Mouse production. The pups will be genotyped by PCR followed by sequencing analysis. Note: Mice homozygous for a knock-out alleles exhibit resistance to induced inflammatory responses. Exon 3 starts from about 100% of the coding region. The knockout of Exon 3~9 will result in frameshift of the gene. The size of intron 2 for 5'-loxP site insertion: 104 bp, and the size of intron 9 for 3'-loxP site insertion: 510 bp. The size of effective cKO region: ~2574 bp. The cKO region does not have any other known gene. Page 1 of 8 https://www.alphaknockout.com Overview of the Targeting Strategy Wildtype allele 5' gRNA region gRNA region 3' 1 2 3 4 5 6 7 8 9 10 1 2 Targeting vector Targeted allele Constitutive KO allele (After Cre recombination) Legends Homology arm Exon of mouse Ripk3 cKO region Exon of mouse Adcy4 loxP site Page 2 of 8 https://www.alphaknockout.com Overview of the Dot Plot Window size: 10 bp Forward Reverse Complement Sequence 12 Note: The sequence of homologous arms and cKO region is aligned with itself to determine if there are tandem repeats.
    [Show full text]
  • RIPK3 Antibody (N-Term) Purified Rabbit Polyclonal Antibody (Pab) Catalog # Ap7184a
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 RIPK3 Antibody (N-term) Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP7184a Specification RIPK3 Antibody (N-term) - Product Information Application WB, IHC-P,E Primary Accession Q9Y572 Reactivity Human Host Rabbit Clonality Polyclonal Isotype Rabbit Ig Antigen Region 1-30 RIPK3 Antibody (N-term) - Additional Information Gene ID 11035 Other Names Western blot analysis of RIP3 (RIPK3) Receptor-interacting Antibody (N-term) (Cat.# AP7184a) in Ramos serine/threonine-protein kinase 3, RIP-like cell line lysates (35ug/lane). RIPK3 (arrow) protein kinase 3, Receptor-interacting was detected using the purified Pab. protein 3, RIP-3, RIPK3, RIP3 Target/Specificity This RIPK3 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 1-30 amino acids from the N-terminal region of human RIPK3. Dilution WB~~1:1000 IHC-P~~1:10~50 Format Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is prepared by Saturated Formalin-fixed and paraffin-embedded Ammonium Sulfate (SAS) precipitation human lung carcinoma tissue reacted with followed by dialysis against PBS. RIP3 (RIPK3) antibody (N-term) (Cat.# AP7184a), which was peroxidase-conjugated Storage to the secondary antibody, followed by DAB Maintain refrigerated at 2-8°C for up to 2 staining. This data demonstrates the use of weeks. For long term storage store at -20°C this antibody for immunohistochemistry; in small aliquots to prevent freeze-thaw clinical relevance has not been evaluated. cycles. Precautions RIPK3 Antibody (N-term) - Background RIPK3 Antibody (N-term) is for research use only and not for use in diagnostic or The product of this gene is a member of the therapeutic procedures.
    [Show full text]
  • RIPK3 Antibody (C-Term) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # Ap14554b
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 RIPK3 Antibody (C-term) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP14554b Specification RIPK3 Antibody (C-term) - Product Information Application WB, IHC-P,E Primary Accession Q9Y572 Other Accession NP_006862.2 Reactivity Human Host Rabbit Clonality Polyclonal Isotype Rabbit Ig Calculated MW 56887 Antigen Region 480-509 RIPK3 Antibody (C-term) - Additional Information Gene ID 11035 Other Names Receptor-interacting Anti-RIPK3 Antibody (C-term) at 1:500 serine/threonine-protein kinase 3, RIP-like dilution + HT-29 whole cell lysate protein kinase 3, Receptor-interacting Lysates/proteins at 20 µg per lane. protein 3, RIP-3, RIPK3, RIP3 Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Target/Specificity Predicted band size : 57 kDa This RIPK3 antibody is generated from Blocking/Dilution buffer: 5% NFDM/TBST. rabbits immunized with a KLH conjugated synthetic peptide between 480-509 amino acids from the C-terminal region of human RIPK3. Dilution WB~~1:1000 IHC-P~~1:10~50 Format Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification. Storage RIPK3 Antibody (C-term) (Cat. #AP14554b) Maintain refrigerated at 2-8°C for up to 2 western blot analysis in K562 cell line lysates weeks. For long term storage store at -20°C (35ug/lane).This demonstrates the RIPK3 in small aliquots to prevent freeze-thaw antibody detected the RIPK3 protein (arrow).
    [Show full text]
  • 1 LUBAC Is Essential for Embryogenesis by Preventing Cell
    1 LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis 2 3 Nieves Peltzer1*, Maurice Darding1*, Antonella Montinaro1, Peter Draber1, Helena Draberova1, 4 Sebastian Kupka1, Eva Rieser1, Amanda Fisher2, Ciaran Hutchinson3, Lucia Taraborrelli1, Torsten 5 Hartwig1, Elodie Lafont1, Tobias L. Haas4, Yutaka Shimizu1, Charlotta Böiers1, Aida Sarr1, James 6 Rickard5, Silvia Alvarez-Diaz5, Michael T. Ashworth3, Allison Beal6, Tariq Enver1, John Bertin6, 7 William Kaiser2, Andreas Strasser5, John Silke5, Philippe Bouillet5 and Henning Walczak1,7 8 9 10 1UCL Cancer Institute, University College London, 72 Huntley Street, London WC1E 6DD, UK 11 2University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA 12 3UCL Great Ormond Street Institute of Child Health, 30 Guilford St, London, UK 13 4Institute of General Pathology, Università Cattolica del Sacro Cuore, 00168, Rome, Italy 14 5The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, 15 Australia 16 6Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, 17 GlaxoSmithKline, Collegeville, Pennsylvania, USA 18 7Correspondence to: Henning Walczak, PhD; E-mail: [email protected]. Phone +44 207 679 6471 19 20 * These authors contributed equally. 21 22 1 23 The Linear Ubiquitin chain Assembly Complex (LUBAC) is required for optimal gene activation 24 and prevention of cell death upon activation of immune receptors, including TNFR11. Deficiency 25 in the LUBAC components SHARPIN or HOIP in mice results in severe inflammation in 26 adulthood or embryonic lethality, respectively, due to deregulation of TNFR1-mediated cell 27 death2-8. In humans, deficiency in the third LUBAC component, HOIL-1, causes autoimmunity 28 and inflammatory disease, similar to HOIP deficiency, whereas HOIL-1 deficiency in mice was 29 reported to cause no overt phenotype9-11.
    [Show full text]