DOCSLIB.ORG

Thesis Reference

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Thesis Reference Thesis Synthetic Activation of Aberrantly Silenced Genes in Diseases HAFNER, Nicolas Abstract DNA methylation influences the accessibility of DNA to transcription factors (TFs). Aberrant hypermethylation and subsequent inhibition of tumor suppressor gene (TSG) promoters is a hallmark in many cancers. Reactivation of TSGs in cancer cells by targeted demethylation of their promoters is an attractive therapeutic approach. A recently identified class of TFs, called super pioneer factors (SPFs), can bind to methylated DNA, leading to demethylation. Here, we explore the possibility of reactivation of methylated promoters of TSGs, by recruitment of SPFs via CRISPR/Cas9-mediated insertion of their binding motifs. To find additional therapeutic targets for epigenome editing, we investigated the epigenetic profile of various adipose tissues. Here, we performed methylome and transcriptome analysis on visceral adipose tissue in mice after and before treatment with cold and antibiotics. Reference HAFNER, Nicolas. Synthetic Activation of Aberrantly Silenced Genes in Diseases. Thèse de doctorat : Univ. Genève, 2020, no. Sc. Vie 49 DOI : 10.13097/archive-ouverte/unige:141767 URN : urn:nbn:ch:unige-1417679 Available at: http://archive-ouverte.unige.ch/unige:141767 Disclaimer: layout of this document may differ from the published version. 1 / 1 UNIVERSITÉ DE GENÈVE FACULTÉ DE MÉDECINE Section de médecine fondamentale Département de médecine génétique et développement Professeur Rabih Murr SYNTHETIC ACTIVATION OF ABERRANTLY SILENCED GENES IN DISEASES THÈSE présentée aux Facultés de médecine et des sciences de l’Université de Genève pour obtenir le grade de Docteur ès sciences en sciences de la vie, mention Sciences biomédicales Par Nicolas Hafner de Mannheim (Allemagne) Thèse N o 49 GENÈVE 2019 IIFE SCIENCES PhD SCHOOL GENEVA DocroRAT Ès scrENcES EN ScTENCES DE LAVTE DES FACUITÉs DE MÉoEcINE ET DES SCIENCES MENTToN ScTENCES BroMÉorcnlrs Thèse de Mr Nicolas HAFNER intitulée: < Synthetic Activation of Aberrantly Silenced Genes in Diseases r> Les Facultés de médecine et des sciences, sur le préavis de Monsieur Rabih MURR, professeur assistant et directeur de thèse (Département de Biologie Moléculaire), Monsieur David SHORE, professeur (Unité des Produits Naturels Bioactifs), Monsieur Mirko TRAJKOVSKI, professeur ordinaire (Département de Physiologie Cellulaire et Métabolisme), Monsieur Zdenko HERCEG (Centre lnternational de Recherche sur le Cancer, Lyon, France) autorisent I'impression de la présente thèse, sans exprimer d'opinion sur les propositions qui y sont énoncées. Genève, le 20 février 2020 Thèse - 49 Le Le Décanat Faculté de médecine Faculté des sciences N.B. - La thèse doit porter la déclaration précédente et remplir les conditions énumérées dans les "lnformations relatives aux thèses de doctorat à I'Université de Genève". a FACULTE DE MEDECINE UNIVERSITE FACULTÉ DEs SCIENCES DE GENEVE Table of Contents Abbreviations ............................................................................................................................ V Acknowledgements ............................................................................................................... VIII Abstract ..................................................................................................................................... X Résumé .................................................................................................................................... XII 1 Introduction ........................................................................................................................ 1 1.1 Epigenetics and the Regulation of Gene Expression ................................................... 1 1.1.1 From Information to Function: Gene Expression and its Regulation .................. 1 1.1.2 The Biology of Transcription Factors .................................................................. 7 1.1.3 Chromatin and Epigenetics ................................................................................ 12 1.1.4 The 3D Architecture of the Genome .................................................................. 15 1.1.5 Accessing Nucleosomal DNA ............................................................................ 19 1.1.6 Histone Modifications ........................................................................................ 27 1.1.7 DNA Methylation ............................................................................................... 32 1.2 CRISPR ..................................................................................................................... 66 1.2.1 Exploits from an Ancient War ........................................................................... 66 1.2.2 Harnessing CRISPR for Genome Editing .......................................................... 72 1.2.3 No Cutting Needed: dCas9-Effectors ................................................................. 75 1.2.4 Opportunities and Challenges ............................................................................ 79 2 Aim of the Thesis ............................................................................................................. 83 3 Results .............................................................................................................................. 85 II 3.1 Reactivation of Epigenetically Silenced Tumor Suppressor Genes with Super Pioneer Transcription Factors .......................................................................................... 85 3.1.1 Validation of the Experimental Approach by Targeting TSGs with dCas9- TET1 87 3.1.2 Phenotypic Changes Subsequent to Targeted p16 Reactivation .................. 92 3.1.3 Insertion of the CTCF motif into TSG Promoter Regions ........................... 96 3.1.4 Insertion of SPF motifs into a Methylation Reporter ................................. 102 3.2 Cell Fate of Adipose Tissues: New Targets for Epigenome Editing ................. 110 3.2.1 Identification of Differentially Expressed Genes and Differentially Methylated Regions Upon Browning in VAT Adipocytes ........................................ 112 3.2.3 Differentiation of 3T3L1-preadipocytes under the Influence of 5-aza ...... 120 4 Discussion ...................................................................................................................... 123 4.1 Experimental Approach ........................................................................................ 123 4.2 Interpretation of the Results ................................................................................. 130 4.3 Outlook and Perspectives ...................................................................................... 133 5 Materials and Methods ................................................................................................... 137 5.1 Cell Culture.............................................................................................................. 137 5.2 Treatment of Dazl-Snrpn-GFP V6.5 mESCs with 5-azacytidine ............................ 137 5.3 Cloning of guide RNAs ........................................................................................... 137 5.4 Lentivirus Production and Transduction ................................................................. 139 5.5 SPF Motif Insertion with CRISPR/Cas9 ................................................................. 141 5.6 Genotyping .............................................................................................................. 143 III 5.7 Bisulfite Conversion, PCR, and Sequencing ........................................................... 144 5.8 RNA Extraction and qPCR ...................................................................................... 144 5.9 Protein Extraction and Western Blot ....................................................................... 146 5.10 Cell Counting Assay ............................................................................................ 147 5.11 Live Cell Imaging ................................................................................................ 148 5.12 Cell Cycle Analysis of Live Cells ........................................................................ 148 5.13 Senescence β-Galactosidase Staining .................................................................. 148 5.14 Flow Cytometry of Dazl-Snrpn Reporter Cells ................................................... 149 5.15 Animal Housing and CAx Treatment .................................................................. 149 5.16 Whole Genome Bisulfite Sequencing .................................................................. 149 5.17 RNA Sequencing ................................................................................................. 150 5.18 Differentiation of 3T3-L1 Pre-Adipocytes .......................................................... 151 6 References ...................................................................................................................... 152 IV Abbreviations 3C Chromosome conformation capture 4C Circular 3C 5-mC 5-methylcytosine A Adenine aa Amino acid AAV Adeno-associated virus BAT Brown adipose tissue bp Base pair BMI Body-mass index BS Bisulfite sequencing C Cytosine Cas CRISPR associated protein CAx Cold/antibiotics treatment CDK Cyclin-dependent kinase cDNA Cyclic DNA CGI CpG island ChIP Chromatin immunoprecipitation Cidea Cell death activator CpG Cytosine followed by a guanine CRE Cis-regulatory
Load more

Recommended publications
  • Datasheet for Histone H2B Human, Recombinant (M2505; Lot 0031404)
    Supplied in: 20 mM Sodium Phosphate (pH 7.0), Quality Control Assays: Ionization-Time of Flight Mass Spectrometry). The Histone H2B 300 mM NaCl and 1 mM EDTA. SDS-PAGE: 0.5 µg, 1.0 µg, 2.0 µg, 5.0 µg, average mass calculated from primary sequence Human, Recombinant 10.0 µg Histone H2B Human, Recombinant were is 13788.97 Da. This confirms the protein identity Note: The protein concentration (1 mg/ml, 73 µM) loaded on a 10–20% Tris-Glycine SDS-PAGE gel as well as the absence of any modifications of the is calculated using the molar extinction coefficient and stained with Coomassie Blue. The calculated histone. 1-800-632-7799 for Histone H2B (6400) and its absorbance at molecular weight is 13788.97 Da. Its apparent [email protected] 280 nm (3,4). 1.0 A units = 2.2 mg/ml N-terminal Protein Sequencing: Protein identity 280 molecular weight on 10–20% Tris-Glycine SDS- www.neb.com was confirmed using Edman Degradation to PAGE gel is ~17 kDa. M2505S 003140416041 Synonyms: Histone H2B/q, Histone H2B.1, sequence the intact protein. Histone H2B-GL105 Mass Spectrometry: The mass of purified Histone H2B Human, Recombinant is 13788.5 Da Protease Assay: After incubation of 10 µg of M2505S B r kDa 1 2 3 4 5 6 7 as determined by ESI-TOF MS (Electrospray Histone H2B Human, Recombinant with a standard 250 4.0 mixture of proteins for 2 hours at 37°C, no 100 µg 1.0 mg/ml Lot: 0031404 150 13788.5 100 proteolytic activity could be detected by SDS- RECOMBINANT Store at –20°C Exp: 4/16 80 PAGE.
    [Show full text]
  • Histone Variants: Deviants?
    Downloaded from genesdev.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW Histone variants: deviants? Rohinton T. Kamakaka2,3 and Sue Biggins1 1Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA; 2UCT/National Institutes of Health, Bethesda, Maryland 20892, USA Histones are a major component of chromatin, the pro- sealing the two turns of DNA. The nucleosome filament tein–DNA complex fundamental to genome packaging, is then folded into a 30-nm fiber mediated in part by function, and regulation. A fraction of histones are non- nucleosome–nucleosome interactions, and this fiber is allelic variants that have specific expression, localiza- probably the template for most nuclear processes. Addi- tion, and species-distribution patterns. Here we discuss tional levels of compaction enable these fibers to be recent progress in understanding how histone variants packaged into the small volume of the nucleus. lead to changes in chromatin structure and dynamics to The packaging of DNA into nucleosomes and chroma- carry out specific functions. In addition, we review his- tin positively or negatively affects all nuclear processes tone variant assembly into chromatin, the structure of in the cell. While nucleosomes have long been viewed as the variant chromatin, and post-translational modifica- stable entities, there is a large body of evidence indicat- tions that occur on the variants. ing that they are highly dynamic (for review, see Ka- makaka 2003), capable of being altered in their compo- Supplemental material is available at http://www.genesdev.org. sition, structure, and location along the DNA. Enzyme Approximately two meters of human diploid DNA are complexes that either post-translationally modify the packaged into the cell’s nucleus with a volume of ∼1000 histones or alter the position and structure of the nucleo- µm3.
    [Show full text]
  • How Human H1 Histone Recognizes DNA
    molecules Article How Human H1 Histone Recognizes DNA Olesya P. Luzhetskaya, Sergey E. Sedykh and Georgy A. Nevinsky * Institute of Chemical Biology and Fundamental Medicine, SD of Russian Academy of Sciences, 8 Lavrentiev Ave., 630090 Novosibirsk, Russia; [email protected] (O.P.L.); [email protected] (S.E.S.) * Correspondence: [email protected]; Tel.: +7-383-363-51-26; Fax: +7-383-363-51-53 Received: 11 August 2020; Accepted: 1 October 2020; Published: 5 October 2020 Abstract: Linker H1 histone is one of the five main histone proteins (H1, H2A, H2B, H3, and H4), which are components of chromatin in eukaryotic cells. Here we have analyzed the patterns of DNA recognition by free H1 histone using a stepwise increase of the ligand complexity method; the affinity of H1 histone for various single- and double-stranded oligonucleotides (d(pN)n; n = 1–20) was evaluated using their competition with 12-mer [32P]labeled oligonucleotide and protein–oligonucleotide complex delaying on nitrocellulose membrane filters. It was shown that minimal ligands of H1 histone (like other DNA-dependent proteins and enzymes) are different mononucleotides (dNMPs; Kd = (1.30 0.2) 2 ± 10 M). An increase in the length of single-stranded (ss) homo- and hetero-oligonucleotides (d(pA)n, × − d(pT)n, d(pC)n, and d(pN)n with different bases) by one nucleotide link regardless of their bases, leads to a monotonic increase in their affinity by a factor of f = 3.0 0.2. This factor f corresponds ± to the Kd value = 1/f characterizing the affinity of one nucleotide of different ss d(pN)n for H1 at n = 2–6 (which are covered by this protein globule) is approximately 0.33 0.02 M.
    [Show full text]
  • The Heterochromatin Condensation State in Peripheral “Gene Poor” and Central “Gene Rich” Nuclear Regions of Less Differe
    L al of euk rn em u i o a J Journal of Leukemia Karel Smetana, J Leuk 2014, 2:4 ISSN: 2329-6917 DOI: 10.4172/2329-6917.1000151 Research Article Open Access The Heterochromatin Condensation State in Peripheral “Gene Poor” and Central “Gene Rich” Nuclear Regions of Less Differentiated and Mature Human Leukemic Cells: A Mini-Review with Additional Original Observations Karel Smetana* Institute of Hematology and Blood Transfusion, Prague, Czech Republic *Corresponding author: Karel Smetana, Senior scientist Institute of Hematology and Blood Transfusion, U nemocnice 1, 128 20 Prague, Czech Republic, Tel: 420 739906473; E-mail: [email protected] Rec date: May 22, 2014; Acc date: Aug 28, 2014; Pub date: Aug 30, 2014 Copyright: © 2014 Karel Smenata. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract In the morphological cytology the heterochromatin is one of very useful tools for the cell identification including the differentiation and maturation stage. However, the heterochromatin condensation state was less studied although it appeared to be different in “gene rich” central and “gene poor” peripheral nuclear regions. The heavy heterochromatin condensation state in the central “gene rich” nuclear regions might reflect a marked structural stability and protect the genomic integrity. It must be also noted that the heterochromatin condensation state in these nuclear regions is more variable than in the nuclear periphery because of the presence of more as well as less condensed heterochromatin territories.
    [Show full text]
  • The Mechanism of Cross-Talk Between Histone H2B Ubiquitination and H3 Methylation by Dot1l
    bioRxiv preprint doi: https://doi.org/10.1101/501098; this version posted December 22, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. The mechanism of cross-talk between histone H2B ubiquitination and H3 methylation by Dot1L Evan J. Worden, Niklas Hoffmann, Chad Hicks and Cynthia Wolberger* Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD USA *Corresponding author: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/501098; this version posted December 22, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract Methylation of histone H3, lysine 79 (H3K79), by Dot1L is a hallmark of actively transcribed genes that depends on monoubiquitination of H2B at lysine 120 (H2B-Ub), and is a well-characterized example of histone modification cross-talk that is conserved from yeast to humans. The mechanism by which H2B-Ub stimulates Dot1L to methylate the relatively inaccessible histone core H3K79 residue is unknown. The 3.0 Å resolution cryo-EM structure of Dot1L bound to ubiquitinated nucleosome reveals that Dot1L contains binding sites for both ubiquitin and the histone H4 tail, which establish two regions of contact that stabilize a catalytically competent state and positions the Dot1L active site over H3K79.
    [Show full text]
  • Histone H1 Is Essential for Mitotic Chromosome Architecture
    Published Online: 20 June, 2005 | Supp Info: http://doi.org/10.1083/jcb.200503031 JCB: ARTICLE Downloaded from jcb.rupress.org on May 12, 2019 Histone H1 is essential for mitotic chromosome architecture and segregation in Xenopus laevis egg extracts Thomas J. Maresca, Benjamin S. Freedman, and Rebecca Heald Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720 uring cell division, condensation and resolution of spindle. Although functional kinetochores assembled, chromosome arms and the assembly of a func- aligned, and exhibited poleward movement, long and D tional kinetochore at the centromere of each sister tangled chromosome arms could not be segregated in chromatid are essential steps for accurate segregation of anaphase. Histone H1 depletion did not significantly affect the genome by the mitotic spindle, yet the contribution of the recruitment of known structural or functional chromo- individual chromatin proteins to these processes is poorly somal components such as condensins or chromokinesins, understood. We have investigated the role of embryonic suggesting that the loss of H1 affects chromosome archi- linker histone H1 during mitosis in Xenopus laevis egg tecture directly. Thus, our results indicate that linker histone extracts. Immunodepletion of histone H1 caused the assem- H1 plays an important role in the structure and function of bly of aberrant elongated chromosomes that extended off vertebrate chromosomes in mitosis. the metaphase plate and outside the perimeter of the Introduction Correct transmission of the genome by the mitotic spindle complexes condensin and cohesin, which are thought to form during cell division requires dramatic changes in chromosome ring structures that generate chromosome super coiling or architecture.
    [Show full text]
  • Epigenetic Control of Mammalian Centromere Protein Binding: Does DNA Methylation Have a Role?
    Journal of Cell Science 109, 2199-2206 (1996) 2199 Printed in Great Britain © The Company of Biologists Limited 1996 JCS3386 Epigenetic control of mammalian centromere protein binding: does DNA methylation have a role? Arthur R. Mitchell*, Peter Jeppesen, Linda Nicol†, Harris Morrison and David Kipling MRC Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK *Author for correspondence (internet [email protected]) †Present address: MRC Reproductive Biology Unit, Edinburgh, UK SUMMARY Chromosome 1 of the inbred mouse strain DBA/2 has a block of minor satellite DNA sequences on chromosome 1. polymorphism associated with the minor satellite DNA at The binding of the CENP-E protein does not appear to be its centromere. The more terminal block of satellite DNA affected by demethylation of the minor satellite sequences. sequences on this chromosome acts as the centromere as We present a model to explain these observations. This shown by the binding of CREST ACA serum, anti-CENP- model may also indicate the mechanism by which the B and anti-CENP-E polyclonal sera. Demethylation of the CENP-B protein recognises specific sites within the arrays minor satellite DNA sequences accomplished by growing of minor satellite DNA on mouse chromosomes. cells in the presence of the drug 5-aza-2′-deoxycytidine results in a redistribution of the CENP-B protein. This protein now binds to an enlarged area on the more terminal Key words: Centromere satellite DNA, Demethylation, Centromere block and in addition it now binds to the more internal antibody INTRODUCTION A common feature of many mammalian pericentromeric domains is that they contain families of repetitive DNA The centromere of mammalian chromosomes is recognised at sequences (Singer, 1982).
    [Show full text]
  • H2A Ubiquitylated Mononucleosomes Next-Generation Substrates for Deubiquitylation Enzyme (DUB) Assays
    H2A Ubiquitylated Mononucleosomes Next-Generation Substrates for Deubiquitylation Enzyme (DUB) Assays Next-Generation DUB Assay Substrates are here. Get results that matter. • Enabling access to DUB targets that require nucleosome substrates in vitro • Proper substrates for DUB inhibitor development • Unmatched quality control for results you can trust Histone monoubiquitylation (ub1) acts as a critical signaling center that regulates cascades of downstream epigenetic enzymes to modify gene transcription. The physiological substrate for chromatin-targeting DUBs is the nucleosome (Nuc), the basic repeating unit of chromatin (comprised of histone proteins wrapped by DNA). Current high-throughput screening (HTS) DUB assays use unnatural modified or diubiquitin conjugates as substrates, which poorly mimic endogenous targets in vivo. In collaboration with Boston Biochem, EpiCypher is delivering ubiquitylated nucleosome substrates for drug screening and chromatin biology research. FIGURE 1 Ub Ub Schematic representation of mononucleosoms assembled from recombinant human histones Ub expressed in E. coli (two each of histones H2A, H2B, H3 and H4). H2A H2A Approximately 50% of the nucleosomes are monoubiquitylated on histone H2A lysine 118, while the other 50% are monoubiquitylated on both histone H2A lysine 118 and histone H2A lysine 119 (multi-mono- ubiquitylated). Next Generation Deubiquitylation Enzyme (DUB) Assay Substrates EpiCypher has developed recombinant mononucleosomes carrying monoubiquitylation on H2A. These ubiquitylated nucleosomes are generated enzymatically using the RING1B/BMI1 ubiquitin ligase complex. The resulting product is highly pure (>95% of nucleosomes are ubiquitylated) and consists of nucleosomes monoubiquitylated at H2A lysine 118/119 (Figure 1; the physiological target of RING1B/BMI1 in vivo). FIGURE 2 Deubiquitylation Assay Data: Mononucleosomes H2A Ubiquityl, Recombinant Human, Biotinylated (1 μg) were employed in a deubiquitylation (DUB) assay using no enzyme (Lane 1), USP5 (Lane 2) or USP16 (Lane 3) and run on an SDS PAGE gel.
    [Show full text]
  • Re-Coding the ‘Corrupt’ Code: CRISPR-Cas9 Interventions in Human Germ Line Editing
    Re-coding the ‘corrupt’ code: CRISPR-Cas9 interventions in human germ line editing CRISPR-Cas9, Germline Intervention, Human Cognition, Human Rights, International Regulation Master Thesis Tilburg University- Law and Technology 2018-19 Tilburg Institute for Law, Technology, and Society (TILT) October 2019 Student: Srishti Tripathy Supervisors: Prof. Dr. Robin Pierce SRN: 2012391 Dr. Emre Bayamlioglu ANR: 659785 Re-coding the ‘corrupt’ code CRISPR-Cas9, Germline Intervention, Human Cognition, Human Rights, International Regulation This page is intentionally left blank 2 Re-coding the ‘corrupt’ code CRISPR-Cas9, Germline Intervention, Human Cognition, Human Rights, International Regulation 3 Re-coding the ‘corrupt’ code CRISPR-Cas9, Germline Intervention, Human Cognition, Human Rights, International Regulation Table of Contents CHAPTER 1: Introduction .............................................................................................................. 6 1.1 Introduction and Review - “I think I’m crazy enough to do it” ......................................................................... 6 1.2 Research Question and Sub Questions .......................................................................................................................... 9 1.4 Methodology ............................................................................................................................................................................. 9 1.4 Thesis structure: .................................................................................................................................................................
    [Show full text]
  • The Basic Karyotype of Rye Giemsa and Fluorescence
    Heredity (1974) 33 (3), 403-408 THEBASIC KARYOTYPE OF RYE (SECALE CEREALE) ANALYSED WITH GIEMSA AND FLUORESCENCE METHODS CANIO G. VOSA Botany School, Oxford Received26.iii.74 SUMMARY The basic karyotype of Rye (Secale cereale L.) has been analysed with several fluorochromes and with a Giemsa staining technique. Only the fluoro- chrome Hoechst 33258 and the Giemsa technique were successful in differenti- ating the heterochromatic segments. Five cultivated varieties were studied. In the haploid set there are 11 large and two small terminally located bands, four small intercalary bands and a variable band adjacent to the nucleolar constriction on chromosome VII which are constant for all varieties. Five more very small intercalary bands occur sporadically in all the varieties. All the chromosomes possess thin centromeric bands. 1. INTRODUCTION RYE is the hardiest of the cereals and its cultivation reaches beyond the Arctic Circle. It is of considerable importance in Russia, Sweden, Poland, the Netherlands and Belgium. In Britain, Rye has ceased to be of any great importance since the change to wheaten bread in the eighteenth century. Compared with other cereals, Rye has fewer varieties and these are not very uniform because of cross-pollination which is the rule in Rye but the exception in other cereal crops. It probably occurred as a weed in fields before being accepted as a crop rather later in the history of civilisation.Its oldest archaeological records date from the first Iron Age (Halstatt period, c. 900-700 B.C.). Thefirst chromosome counts are those of Nemec (1910) as cited by Emme (1928), giving 18 chromosomes for endosperm cells.
    [Show full text]
  • Extra Euchromatic Band in the Qh Region of Chromosome 9
    J Med Genet: first published as 10.1136/jmg.22.2.156 on 1 April 1985. Downloaded from 156 Short reports two centromeres indicated by two distinct C bands but only I HANCKE AND K MILLER one primary constriction at the proximal C band. The two Department of Human Genetics, C bands were separated by chromosomal material staining Medizinische Hochschule Hannover, pale in G banding and intensely dark in R banding (fig 1). Hannover, Both NORs could be observed in satellite associations (fig Federal Republic of Germany. 2). The chromosome was therefore defined as pseudo- dicentric chromosome 21 (pseudic 21). The same chromo- References some was found in the proband's father and paternal Balicek P, Zizka, J. Intercalar satellites of human acrocentric grandmother. chromosomes as a cytological manifestation of polymorphisms Acrocentric chromosomes with a short arm morphology in GC-rich material? Hum Genet 1980;54:343-7. similar to that presented here have been reported by 2 Ing PS, Smith SD. Cytogenetic studies of a patient with Balicek and Zizka.' These authors paid no attention to the mosaicism of isochromosome 13q and a dicentric (Y;13) activity of the centromeres. The suppression of additional translocation showing differential centromeric activity. Clin centromeres is indicated by the presence of only one Genet 1983;24:194-9. primary constriction as shown by Ing and Smith2 in a 3 Passarge E. Analysis of chromosomes in mitosis and evaluation dicentric (Y;13) transtocation. Variants of acrocentric of cytogenetic data. 5. Variability of the karyotype. In: Schwarzacher HG, Wolf U, eds. Methods in human cytogene- chromosomes are often observed in patients with congen- tics.
    [Show full text]
  • Cellular and Molecular Signatures in the Disease Tissue of Early
    Cellular and Molecular Signatures in the Disease Tissue of Early Rheumatoid Arthritis Stratify Clinical Response to csDMARD-Therapy and Predict Radiographic Progression Frances Humby1,* Myles Lewis1,* Nandhini Ramamoorthi2, Jason Hackney3, Michael Barnes1, Michele Bombardieri1, Francesca Setiadi2, Stephen Kelly1, Fabiola Bene1, Maria di Cicco1, Sudeh Riahi1, Vidalba Rocher-Ros1, Nora Ng1, Ilias Lazorou1, Rebecca E. Hands1, Desiree van der Heijde4, Robert Landewé5, Annette van der Helm-van Mil4, Alberto Cauli6, Iain B. McInnes7, Christopher D. Buckley8, Ernest Choy9, Peter Taylor10, Michael J. Townsend2 & Costantino Pitzalis1 1Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK. Departments of 2Biomarker Discovery OMNI, 3Bioinformatics and Computational Biology, Genentech Research and Early Development, South San Francisco, California 94080 USA 4Department of Rheumatology, Leiden University Medical Center, The Netherlands 5Department of Clinical Immunology & Rheumatology, Amsterdam Rheumatology & Immunology Center, Amsterdam, The Netherlands 6Rheumatology Unit, Department of Medical Sciences, Policlinico of the University of Cagliari, Cagliari, Italy 7Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow G12 8TA, UK 8Rheumatology Research Group, Institute of Inflammation and Ageing (IIA), University of Birmingham, Birmingham B15 2WB, UK 9Institute of
    [Show full text]