DOCSLIB.ORG

Chip-Seq Analysis of AIRE, FEZF2, and DEAF1 Transcription Factors in Human Thymus Tissue

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Chip-Seq Analysis of AIRE, FEZF2, and DEAF1 Transcription Factors in Human Thymus Tissue Master’s Thesis 2020 60 ECTS Faculty of Chemistry, Biotechnology, and Food Science (KBM) ChIP-seq analysis of AIRE, FEZF2, and DEAF1 transcription factors in human thymus tissue Dina Ruud Aronsen Biotechnology – molecular biology ChIP-seq analysis of AIRE, FEZF2 and DEAF1 transcription factors in human thymus tissue Dina Ruud Aronsen Supervisors: Prof. Dr. Benedicte A. Lie (Main supervisor) Dr. Teodora Ribarska Assoc. Prof. Siv Kjølsrud Bøhn Master thesis Department of Medical Genetics, Oslo University Hospital Faculty of Chemistry, Biotechnology and Food science Norwegian University of Life Sciences June 2020 i ii ©Dina Ruud Aronsen 2020 ChIP-seq analysis of AIRE, FEZF2 and DEAF1 transcription factors in human thymus tissue https://nmbu.brage.unit.no iii Acknowledgements The work of this thesis was carried out in collaboration with the Department of Medical Genetics, at Oslo University Hospital (OUS), as part of a Master´s Degree in Biotechnology at the Norwegian University of Life Sciences (NMBU), the main affiliation being the Faculty of Chemistry, Biotechnology and Food Science (KBM), from August 2019 to June 2020. Foremost, I would like to thank my main supervisor Benedicte A. Lie for the opportunity to be a part of her thymus group and conduct such interesting research. I am grateful for her guidance, insightful advice and the enthusiasm she has shown for my work. I am thankful for the outstanding work of my co-supervisor Teodora Ribarska, Postdoc. She guided and encouraged me through both the writing process and laboratory work. Her invaluable advice and knowledge have greatly contributed to my understanding throughout this process. Additionally, her laboratory expertise and guidance have helped me improve my laboratory skills and expanded my expertise by introducing me to several laboratory techniques. Additionally, I would like to thank my supervisor at NMBU, Siv Kjølsrud Bøhn, for valuable feedback and guidance. I would like to thank all past and present members of the Immgen-group for welcoming and including me and for providing advice along the way. I would like to extend an extra thank to the thymus-group for advice and for helping with laboratory work. Lastly, I would like to thank my loved ones, who have supported me throughout the entire process. I will be grateful forever for your love and encouragement. Oslo, June 2020 Dina Ruud Aronsen iv Abstract Autoimmune diseases develop as a consequence of incorrect establishment of self-tolerance. A key element in this process is the presentation of tissue-restricted antigens (TRA) by medullary thymic epithelial cells (mTECs) and antigen presenting cells (APCs) to the developing T-cells in the thymus. TRA expression is mostly driven by the transcription factors Autoimmune regulator (AIRE) and Forebrain Embryonic Zinc Finger-Like Protein 2 (FEZF2) in mTECs. Deformed Epidermal Autoregulatory Factor 1 Homolog (DEAF1), present in all thymic APCs, is suggested to also control TRA expression as it does in the lymph node. The purpose of this study was to establish and optimise chromatin immunoprecipitation (ChIP) method for use in human freshly frozen thymus tissues and apply it to determine the genomic binding sites of FEZF2, AIRE and DEAF1 in the infant human thymus in order to better understand the molecular mechanism of T-cell self-tolerance establishment. Aiming to reach the highest possible sensitivity and specificity of the enrichment, we optimised the ChIP procedure using homogenised frozen human thymic tissue (n=3) by testing the parameters: fixation (single or double crosslinking), fragmentation by sonication, antibody type and amount, and immunoprecipitation conditions. The optimal ChIP conditions were found to be: 1) Fixation for 5 min with 1 % formaldehyde and 2.5 mM disuccinimidyl glutarate with 20-30 mg tissue powder ; 2) Sonication to 300-500 bp average DNA fragment size; 3) IP with 5 µg anti-AIRE (GeneTex), 5 µg anti-DEAF1 (LSBio), and 1 µg FEZF2 (Abcam) per 100 µl chromatin (out of 2 mL chromatin isolated from 20-30 mg tissue powder) using 30 µl protein G magnetic beads (Dynabeads) per IP reaction. These condition provided the highest specific recovery of target regions for the three transcription factors, measured by qPCR. The optimal conditions were used in a pilot ChIP-seq experiment to locate binding sites of the transcription factors. Analysis of AIRE, FEZF2, and DEAF1 ChIP-seq data from human thymus and qPCR of ChIP using TEC and APC enriched cell sample isolated from half of a thymus showed low enrichment of target genes, which may be caused by scarcity of cells expressing AIRE, FEZF2, and DEAF1 in bulk thymus tissue or enriched for mTEC and APC thymus cell suspension. To assess the function of AIRE, FEZF2, and DEAF1 in mTECs and thymic APCs, further exploration using CUT&Tag or ChIP-seq will be needed using mTEC and APC enriched cell solution. v vi Table of Contents Acknowledgements ......................................................................................................................... iv Abstract ............................................................................................................................................ v Abbreviations .................................................................................................................................. ix List of Figures ................................................................................................................................. xi 1 Introduction ............................................................................................................................... 1 1.1 The Immune System .......................................................................................................... 1 1.1.1 Cells of the Immune System ...................................................................................... 1 1.1.2 The Innate Immune System ........................................................................................ 5 1.1.3 Human Leukocyte Antigen ........................................................................................ 5 1.1.4 The Adaptive Immune System ................................................................................... 6 1.2 T-cell Development ........................................................................................................... 8 1.2.1 Positive Selection ..................................................................................................... 10 1.2.2 Negative Selection .................................................................................................... 11 1.2.3 Peripheral tolerance .................................................................................................. 13 1.2.4 Antigen-Presenting Cells in the Thymus ................................................................. 13 1.3 AIRE, FEZF2, and DEAF1 Functions in APCs .............................................................. 14 1.3.1 AIRE ......................................................................................................................... 14 1.3.2 FEZF2 ....................................................................................................................... 15 1.3.3 DEAF1 ..................................................................................................................... 17 1.4 Autoimmune Diseases ..................................................................................................... 18 2 Aims ........................................................................................................................................ 20 3 Materials and Methods ............................................................................................................ 21 3.1 Thymus collection ........................................................................................................... 21 3.2 Frozen thymus tissue preparation .................................................................................... 21 3.3 Fresh thymus tissue dissociation ..................................................................................... 21 3.4 Chromatin Immunoprecipitation followed by Sequencing ............................................. 23 3.4.1 Chromatin isolation .................................................................................................. 25 3.4.2 Chromatin Immunoprecipitation .............................................................................. 27 3.4.3 Quantitative PCR (qPCR) ........................................................................................ 28 3.4.4 Library preparation and sequencing ......................................................................... 31 vii 3.5 ChIP DNA analysis ......................................................................................................... 32 3.6 CUT&Tag ........................................................................................................................ 32 4 Results ..................................................................................................................................... 36 4.1 AIRE, FEZF2, and DEAF1 expression in thymus .......................................................... 36 4.2 Chromatin Immunoprecipitation Optimisation ............................................................... 37 4.2.1 Crosslinking and Sonication Optimisation ............................................................... 37 4.2.2 Chromatin Immunoprecipitation
Load more

Recommended publications
  • An Amplified Fatty Acid-Binding Protein Gene Cluster In
    cancers Review An Amplified Fatty Acid-Binding Protein Gene Cluster in Prostate Cancer: Emerging Roles in Lipid Metabolism and Metastasis Rong-Zong Liu and Roseline Godbout * Department of Oncology, Cross Cancer Institute, University of Alberta, Edmonton, AB T6G 1Z2, Canada; [email protected] * Correspondence: [email protected]; Tel.: +1-780-432-8901 Received: 6 November 2020; Accepted: 16 December 2020; Published: 18 December 2020 Simple Summary: Prostate cancer is the second most common cancer in men. In many cases, prostate cancer grows very slowly and remains confined to the prostate. These localized cancers can usually be cured. However, prostate cancer can also metastasize to other organs of the body, which often results in death of the patient. We found that a cluster of genes involved in accumulation and utilization of fats exists in multiple copies and is expressed at much higher levels in metastatic prostate cancer compared to localized disease. These genes, called fatty acid-binding protein (or FABP) genes, individually and collectively, promote properties associated with prostate cancer metastasis. We propose that levels of these FABP genes may serve as an indicator of prostate cancer aggressiveness, and that inhibiting the action of FABP genes may provide a new approach to prevent and/or treat metastatic prostate cancer. Abstract: Treatment for early stage and localized prostate cancer (PCa) is highly effective. Patient survival, however, drops dramatically upon metastasis due to drug resistance and cancer recurrence. The molecular mechanisms underlying PCa metastasis are complex and remain unclear. It is therefore crucial to decipher the key genetic alterations and relevant molecular pathways driving PCa metastatic progression so that predictive biomarkers and precise therapeutic targets can be developed.
    [Show full text]
  • Gastroschisis; Perinatal and Postnatal Aspects
    Gastroschisis; perinatal and postnatal aspects Claar Lap Cover design: Marjorie Lap-Streur Printed by: Gildeprint, Enschede, the Netherlands ISBN: 978-90-393-6684-4 © C.C.M.M. Lap, Utrecht, the Netherlands, 2016 All rights reserved. No part of this thesis may be reproduced or transmitted in any form or by any means without prior permission of the author. The study in Chapter 4 was supported by “vrienden WKZ” The author gratefully acknowledges financial support for printing this thesis by Divison Woman & Baby, University Medical Center Utrecht, ACE Pharmaceuticals, BMA B.V. (Mosos), Olympus Nederland B.V., Memidis Pharma B.V. and ChipSoft. Gastroschisis; perinatal and postnatal aspects Gastroschisis; perinatale en postnatale aspecten (met een samenvatting in het Nederlands) Proefschrift ter verkrijging van de graad van doctor aan de Universiteit Utrecht op gezag van de rector magnificus, prof.dr. G.J. van der Zwaan, ingevolge het besluit van het college voor promoties in het openbaar te verdedigen op donderdag 8 december 2016 des middags te 12.45 uur door Chiara Carolina Monique Maria van Haersma Buma-Lap geboren op 24 november 1982 te Sint-Oedenrode Promotor: Prof. dr. G.H.A. Visser Copromotoren: Dr. G.T.R. Manten Dr. L.R. Pistorius CONTENTS List of Abbreviations 9 Chapter 1 General Introduction 13 PART I: OUTCOME OF GASTROSCHISIS Chapter 2 Outcome of isolated gastroschisis; an international study, systematic 33 review and meta-analysis (Early Hum Dev. 2016;103:209-218) Chapter 3 Functional outcome at school age of children born with
    [Show full text]
  • Vertebrate Fatty Acid and Retinoid Binding Protein Genes and Proteins: Evidence for Ancient and Recent Gene Duplication Events
    In: Advances in Genetics Research. Volume 11 ISBN: 978-1-62948-744-1 Editor: Kevin V. Urbano © 2014 Nova Science Publishers, Inc. Chapter 7 Vertebrate Fatty Acid and Retinoid Binding Protein Genes and Proteins: Evidence for Ancient and Recent Gene Duplication Events Roger S. Holmes Eskitis Institute for Drug Discovery and School of Biomolecular and Physical Sciences, Griffith University, Nathan, QLD, Australia Abstract Fatty acid binding proteins (FABP) and retinoid binding proteins (RBP) are members of a family of small, highly conserved cytoplasmic proteins that function in binding and facilitating the cellular uptake of fatty acids, retinoids and other hydrophobic compounds. Several human FABP-like genes are expressed in the body: FABP1 (liver); FABP2 (intestine); FABP3 (heart and skeletal muscle); FABP4 (adipocyte); FABP5 (epidermis); FABP6 (ileum); FABP7 (brain); FABP8 (nervous system); FABP9 (testis); and FABP12 (retina and testis). A related gene (FABP10) is expressed in lower vertebrate liver and other tissues. Four RBP genes are expressed in human tissues: RBP1 (many tissues); RBP2 (small intestine epithelium); RBP5 (kidney and liver); and RBP7 (kidney and heart). Comparative FABP and RBP amino acid sequences and structures and gene locations were examined using data from several vertebrate genome projects. Sequence alignments, key amino acid residues and conserved predicted secondary and tertiary structures were also studied, including lipid binding regions. Vertebrate FABP- and RBP- like genes usually contained 4 coding exons in conserved locations, supporting a common evolutionary origin for these genes. Phylogenetic analyses examined the relationships and evolutionary origins of these genes, suggesting division into three FABP gene classes: 1: FABP1, FABP6 and FABP10; 2: FABP2; and 3, with 2 groups: 3A: FABP4, FABP8, FABP9 and FABP12; and 3B: and FABP3, FABP5 and FABP7.
    [Show full text]
  • Azadirachtin-A from Azadirachta Indica Impacts Multiple Biological
    This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. Article Cite This: ACS Omega 2019, 4, 9531−9541 http://pubs.acs.org/journal/acsodf Azadirachtin‑A from Azadirachta indica Impacts Multiple Biological Targets in Cotton Bollworm Helicoverpa armigera † ⊥ § ∥ † ⊥ ⊥ Vishal V. Dawkar,*, , Sagar H. Barage, , Ranjit S. Barbole, Amol Fatangare, Susana Grimalt, ‡ # † ‡ ⊥ Saikat Haldar, David G. Heckel, Vidya S. Gupta, Hirekodathakallu V. Thulasiram, AlešSvatos,̌ † and Ashok P. Giri † ‡ Plant Molecular Biology Unit, Division of Biochemical Sciences and Division of Organic Chemistry, CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pune 411008, Maharashtra, India § Bioinformatics Centre, Savitribai Phule Pune University, Ganeshkhind Road, Pune 411007, Maharashtra, India ∥ Amity Institute of Biotechnology (AIB), Amity University, Mumbai−Pune Expressway, Bhatan, Post-Somathne, Panvel, Mumbai 410206, Maharashtra, India ⊥ # Research Group, Mass Spectrometry/Proteomics and Department of Entomology, Max Planck Institute for Chemical Ecology, 07745 Jena, Germany *S Supporting Information ABSTRACT: Azadirachtin-A (AzaA) from the Indian neem tree (Azadirachta indica) has insecticidal properties; however, its molecular mechanism remains elusive. The “targeted and nontargeted proteomic profiling”, metabolomics, matrix- assisted laser desorption/ionization time of flight (MALDI- TOF) imaging, gene expression, and in silico analysis provided clues about its action on Helicoverpa armigera. Fourth instar H. armigera larvae fed on AzaA-based diet (AzaD) suffered from significant mortality, growth retarda- tion, reduced larval mass, complications in molting, and prolonged development. Furthermore, death of AzaD-fed larvae was observed with various phenotypes like bursting, blackening, and half-molting.
    [Show full text]
  • POSTER PRESENTATIONS July 19-21, 2009 Sunday - Tuesday 7:30 A.M.-10:00 A.M
    POSTER PRESENTATIONS July 19-21, 2009 Sunday - Tuesday 7:30 a.m.-10:00 a.m. Exhibit Hall C, Second Floor, DLLCC Cloning Hormone Levels and Identification of Differentially Ex­ pressed Ovarian Genes. Yoji Yamamoto, J. Adam Luck- Downloaded from https://academic.oup.com/biolreprod/article/81/Suppl_2/28/2798222 by guest on 27 September 2021 204. Latrunculin A Dramatically Improves the Developmental enbach, Frederick W. Goetz, Graham Young, Penny Capacity of Nuclear Transfer Embryos Derived from Gene- Swanson. Modified Clawn Miniature Pig Cells. Takehiro Himaki, Hironori Mori, Yamato Mizobe, Kazuchika Miyoshi, Masa- hiro Sato, Sonsin Takao, Mitsutoshi Yoshida. 205. Trichostatin A Treatment Improves the Reprogramability Conservation of Donor Cells from the Haematopoietic Lineage by Somatic Cell Nuclear Transfer. Li-Ying Sung, Chih-Jen Lin, Jie Xu, 213. Recovery of Genetic Diversity Through Xenografting: Sadie L. Marjani, Hongmei Shen, Hui Yu, Tao Cheng, Fine Tuning of This Rescue Technique for Endangered Felids. Xiangzhong Yang, Tian Cindy Tian. Paula Mota, Jens Ehmcke, Kathrin Gassei, Joao Ramalho- Santos, Stefan Schlatt. 206. Global Gene Expression Analysis of Mouse Single Cloned Blastocysts. Atsushi Fukuda, Shinnosuke Morita, Cao 214. Validation of an Anti-Mullerian Hormone (AMH)- Feng, Kimiko Inoue, Narumi Ogonuki, Atsuo Ogura, Yusuke ELISA for Use in an Endangered Marine Mammal, the West Sotomaru, Tomohiro Kono. Indian Manatee (Trichechus manatus). William E. Roude- bush, Dana L. Wetzel, Kevin F. Breuel, John E. Reynolds. 207. Study of Amino Acid Uptake by Cloned Mouse Embryos. Santhi Potireddy, Zhiming Han, Jay Baltz, Keith Latham. 215. Validation of an Inhibin-B ELISA for Use in an Endangered Marine Mammal, the West Indian Manatee (Trichechus manatus).
    [Show full text]
  • Identification De Facteurs De Régulation Du VIH-1 Chez Les Macrophages Humains
    Identification de facteurs de régulation du VIH-1 chez les macrophages humains Mémoire Yann Breton Maîtrise en microbiologie-immunologie Maître ès sciences (M. Sc.) Québec, Canada © Yann Breton, 2016 Identification de facteurs de régulation du VIH-1 chez les macrophages humains Mémoire Yann Breton Sous la direction de : Michel J. Tremblay, directeur de recherche Résumé Lors d'une exposition au VIH-1, bien qu'une seule petite proportion des macrophages soit infectée, il est proposé que ces cellules jouent un rôle important dans l'infection et la propagation du VIH-1. Pour approfondir nos connaissances dans ce domaine, des analyses transcriptomiques et protéomiques ont été effectuées afin de comparer les MDMs (Macrophages Dérivés de Monocytes) infectés aux non infectés. Ces analyses ont mené à la sélection de 50 gènes dont l'expression est modulée chez les cellules infectées pour effectuer un criblage par siRNA pour leurs rôles fonctionnels dans le cycle viral. Huit cibles ont été identifiées comme des régulateurs de l'infection chez les MDMs, mais seulement le gène MDM2 agissait comme un facteur de susceptibilité. Ce gène a donc été l'objet d'études plus approfondies. L'inhibition de l'expression de MDM2 induit une diminution de moitié de l'expression virale. Nos résultats indiquent que la résistance accrue au VIH-1 associée à l’interférence de MDM2 est maintenue même si le niveau d'ARNm est rétabli, suggérant que cette protéine serait impliquée indirectement dans l'infection par le VIH-1. L'identification des cofacteurs viraux régulés par MDM2 mènera à une compréhension des évènements signalétiques contrôlant la réplication du VIH-1 dans les macrophages.
    [Show full text]
  • The Concise Guide to Pharmacology 2019/20: Introduction and Other Protein Targets
    Alexander, S. P. H., Kelly, E., Mathie, A., Peters, J. A., Veale, E. L., Armstrong, J. F., Faccenda, E., Harding, S. D., Pawson, A. J., Sharman, J. L., Southan, C., Buneman, O. P., Cidlowski, J. A., Christopoulos, A., Davenport, A. P., Fabbro, D., Spedding, M., Striessnig, J., Davies, J. A., & CGTP Collaborators (2019). The Concise Guide to Pharmacology 2019/20: Introduction and Other Protein Targets. British Journal of Pharmacology, 176(S1), S1-S20. https://doi.org/10.1111/bph.14747 Publisher's PDF, also known as Version of record License (if available): CC BY Link to published version (if available): 10.1111/bph.14747 Link to publication record in Explore Bristol Research PDF-document This is the final published version of the article (version of record). It first appeared online via Wiley at https://bpspubs.onlinelibrary.wiley.com/doi/full/10.1111/bph.14747. Please refer to any applicable terms of use of the publisher. University of Bristol - Explore Bristol Research General rights This document is made available in accordance with publisher policies. Please cite only the published version using the reference above. Full terms of use are available: http://www.bristol.ac.uk/red/research-policy/pure/user-guides/ebr-terms/ S.P.H. Alexander et al. The Concise Guide to PHARMACOLOGY 2019/20: Introduction andOther Protein Targets.British Journal of Pharmacology (2019) 176, S1–S20 THE CONCISE GUIDE TO PHARMACOLOGY 2019/20: Introduction and Other Protein Targets Stephen PH Alexander1 , Eamonn Kelly2, Alistair Mathie3 ,JohnAPeters4 , Emma L Veale3
    [Show full text]
  • Characterisation of the P19CL6 Cardiovascular Cell-Line at the Gene Expression Level; Effects of Long-Chain Fatty Acids
    Characterisation of the P19CL6 cardiovascular cell-line at the gene expression level; effects of long-chain fatty acids A thesis submitted in accordance with the requirements of the University of Surrey for the degree of Doctor of Philosophy by Iraj Khodadadi Kohlan September 2005 Centre of Nutrition and Food Safety School of Biomedical and Molecular Sciences University of Surrey Guildford Surrey GU2 7XH ProQuest Number: 27610157 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a com plete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 27610157 Published by ProQuest LLO (2019). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C ode Microform Edition © ProQuest LLO. ProQuest LLO. 789 East Eisenhower Parkway P.Q. Box 1346 Ann Arbor, Ml 48106- 1346 Dedication This thesis is dedicated to my parents and my wife for their enormous support, and to Alisa and Parita 11 Acknowledgment I would like to thank my two supervisors: Dr Bruce Griffin and Dr Alfred Thumser for helping me to conduct this research in such an interesting and rapidly expanding field of science. I have been fortunate to use an excellent selection of techniques in this study. I am honoured for having worked with Dr Bruce Griffin, who has been an encouraging supervisor and great support during my study.
    [Show full text]
  • Widespread Transcriptional Scanning in the Testis Modulates Gene Evolution 2 Rates
    bioRxiv preprint doi: https://doi.org/10.1101/282129; this version posted January 22, 2019. 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. Widespread transcriptional scanning in the testis modulates gene evolution 2 rates 4 Bo Xia1, Yun Yan1, Maayan Baron1, Florian Wagner1, Dalia Barkley1, Marta Chiodin1, Sang Y. Kim5, David L. Keefe3, Joseph P. Alukal3, Jef D. Boeke2,4 and Itai Yanai1,2* 6 1 Institute for Computational Medicine, NYU Langone Health, New York, NY 10016, USA 2 Department of Biochemistry and Molecular Pharmacology, NYU Langone Health, New York, NY 8 10016, USA 3 Department of Obstetrics and Gynecology, NYU Langone Health, New York, NY 10016, USA 10 4 Institute for Systems Genetics, NYU Langone Health, New York, NY 10016, USA 5 Department of Pathology, NYU Langone Health, New York, NY 10016, USA 12 *Correspondence to: [email protected] 14 Abstract 16 The testis expresses the largest number of genes of any mammalian organ, a finding that has long puzzled molecular biologists. Analyzing our single cell transcriptomic maps of human and mouse 18 spermatogenesis, we provide evidence that this widespread transcription serves to maintain DNA sequence integrity in the male germline by correcting DNA damage through “transcriptional scanning”. 20 Supporting this model, we find that genes expressed during spermatogenesis display lower mutation rates on the transcribed strand and have low diversity in the population.
    [Show full text]
  • INTRODUCING a NOVEL METHOD for GENETIC ANALYSIS of AUTISM SPECTRUM DISORDER Sepideh Nouri
    The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of The University of Texas MD Anderson Cancer Biomedical Sciences Dissertations and Theses Center UTHealth Graduate School of (Open Access) Biomedical Sciences 12-2013 INTRODUCING A NOVEL METHOD FOR GENETIC ANALYSIS OF AUTISM SPECTRUM DISORDER sepideh nouri Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Bioinformatics Commons, Computational Biology Commons, and the Genomics Commons Recommended Citation nouri, sepideh, "INTRODUCING A NOVEL METHOD FOR GENETIC ANALYSIS OF AUTISM SPECTRUM DISORDER" (2013). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 417. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/417 This Thesis (MS) is brought to you for free and open access by the The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences at DigitalCommons@TMC. It has been accepted for inclusion in The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) by an authorized administrator of DigitalCommons@TMC. For more information, please contact [email protected]. INTRODUCING A NOVEL METHOD FOR GENETIC ANALYSIS OF AUTISM SPECTRUM DISORDER by Sepideh Nouri, M.S. APPROVED: Eric Boerwinkle, Ph.D., Supervisor Kim-Anh Do, Ph.D. Alanna Morrison, Ph.D. James Hixson, Ph.D. Paul Scheet, Ph.D. APPROVED: Dean, The University of Texas Graduate School of Biomedical Sciences at Houston INTRODUCING A NOVEL METHOD FOR GENETIC ANALYSIS OF AUTISM SPECTRUM DISORDER A THESIS Presented to the Faculty of The University of Texas Health Science Center at Houston and The University of Texas M.
    [Show full text]
  • Y Chromosomal Noncoding RNA Regulates Autosomal Gene Expression Via Pirnas in Mouse Testis Hemakumar M
    bioRxiv preprint doi: https://doi.org/10.1101/285429; this version posted March 24, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Y chromosomal noncoding RNA regulates autosomal gene expression via piRNAs in mouse testis Hemakumar M. Reddy,1,2,11 Rupa Bhattacharya,1,3,11 Zeenath Jehan,1,4 Kankadeb Mishra,1,5 Pranatharthi Annapurna,1,6 Shrish Tiwari,1 Nissankararao Mary Praveena,1 Jomini Liza Alex,1 Vishnu M Dhople,1,7 Lalji Singh,10 Mahadevan Sivaramakrishnan,1,8 Anurag Chaturvedi,1,9 Nandini Rangaraj1, Shiju Michael Thomas1, Badanapuram Sridevi,1 Sachin Kumar,1 Ram Reddy Dereddi1, Sunayana Rayabandla1, Rachel A. Jesudasan1* 1Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad, Telengana – 500007, India. Present address: 2Division of Paediatric Neurology, Department of Paediatrics, University of Florida College of Medicine, Gainesville, FL, USA 3Lawrenceville, New Jersey 08648, USA 4Department of Genetics and Molecular Medicines, Vasavi Medical and Research Centre, 6-1-91 Khairatabad, Hyderabad 500 004 India. 5Department of Medical and Clinical Genetics, Goteborg University, Goteborg, Sweden 6National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India 7Ernst-Moritz-Arndt-University of Greifswald Interfaculty Institute for Genetics and Functional Genomics, Department of Functional Genomics, Friedrich-Ludwig-Jahn- Straße 15 a, 17487 Greifswald, Germany 8Jubilant Biosystems Ltd., #96, Industrial Suburb, 2nd Stage, Yeshwantpur, Bangalore- 560022, Karnataka, India 9Laboratory of Biodiversity and Evolutionary Genomics, Charles Deberiotstraat, Leuven, Belgium. 10Deceased. 11These authors contributed equally to the work *Correspondence: Email ID: [email protected]; [email protected] 0 bioRxiv preprint doi: https://doi.org/10.1101/285429; this version posted March 24, 2018.
    [Show full text]
  • Structure of the Human-Heart Fatty-Acid-Binding Protein 3 In
    diffraction structural biology Journal of Synchrotron Structure of the human-heart fatty-acid-binding Radiation protein 3 in complex with the fluorescent probe ISSN 0909-0495 1-anilinonaphthalene-8-sulphonic acid Received 28 May 2013 a,b a,b a,c a,c Accepted 30 July 2013 Mika Hirose, Shigeru Sugiyama, * Hanako Ishida, Mayumi Niiyama, Daisuke Matsuoka,a,b Toshiaki Hara,a,b Eiichi Mizohata,c Satoshi Murakami,d Tsuyoshi Inoue,c Shigeru Matsuokaa,b and Michio Murataa,b PDB Reference: 3wbg aJST, ERATO, Lipid Active Structure Project, 1-1 Machikaneyama-cho, Toyonaka 560-0043, Japan, bGraduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka 560-0043, Japan, cGraduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan, and dGraduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama, Kanagaw 226-8501, Japan. E-mail: [email protected] Heart-type fatty-acid-binding protein (FABP3), which is a cytosolic protein abundantly found in cardiomyocytes, plays a role in trafficking fatty acids throughout cellular compartments by reversibly binding intracellular fatty acids with relatively high affinity. The fluorescent probe 1-anilinonaphthalene-8- sulfonate (ANS) is extensively utilized for examining the interaction of ligands with fatty-acid-binding proteins. The X-ray structure of FABP3 was determined in the presence of ANS and revealed the detailed ANS-binding mechanism. Furthermore, four water molecules were clearly identified in the binding cavity. Through these water molecules, the bound ANS molecule forms indirect hydrogen-bond interactions with FABP3. The adipocyte-type fatty-acid-binding protein (FABP4) exhibits 67% sequence identity with FABP3 and its crystal structure is almost the same as that of FABP3.
    [Show full text]