DOCSLIB.ORG

Development of Transgenic Livestock with Reduced Myostatin

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Development of Transgenic Livestock with Reduced Myostatin DEVELOPMENT OF TRANSGENIC LIVESTOCK WITH REDUCED MYOSTATIN EXPRESSION USING RNA INTERFERENCE A Dissertation by KIMBERLY JEAN TESSANNE Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY December 2009 Major Subject: Veterinary Physiology DEVELOPMENT OF TRANSGENIC LIVESTOCK WITH REDUCED MYOSTATIN EXPRESSION USING RNA INTERFERENCE A Dissertation by KIMBERLY JEAN TESSANNE Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Approved by: Chair of Committee, Mark Westhusin Committee Members, Charles Long Thomas Spencer Nancy Ing Head of Department, Glen Laine December 2009 Major Subject: Veterinary Physiology iii ABSTRACT Development of Transgenic Livestock with Reduced Myostatin Expression Using RNA Interference. (December 2009) Kimberly Jean Tessanne, B.S., The Ohio State University; M.S., University of Wisconsin-Madison Chair of Advisory Committee: Dr. Mark Westhusin RNA interference (RNAi) is a means of regulating gene expression by targeting mRNA in a sequence-specific manner for degradation or translational inhibition. Short hairpin RNAs (shRNAs) and short interfering RNAs (siRNAs) have been extensively employed for manipulating gene expression in a wide range of species. The goal for this research was to produce transgenic livestock in which myostatin, a negative regulator of muscle growth, has been targeted for silencing by RNAi. This would demonstrate the utility of RNAi for reducing gene expression in large animal species. To successfully target the myostatin gene for reduction, siRNAs were designed to target the both the bovine and caprine myostatin mRNA sequence. These were then tested for effectiveness in vitro using both an HEK 293T cell line expressing caprine myostatin as well as adult bovine muscle cells. The most effective siRNA, GDF8-1026, was cloned into a lentiviral plasmid and used to transduce bovine fetal fibroblasts for somatic cell nuclear transfer cloning as well as perivitelline injection of in vitro produced bovine embryos. To date, seven pregnancies have been established using these two methods. iv Concern over the possibility of off-target effects associated with the expression of shRNAs in vivo prompted investigation into tissue-specific expression. Therefore, investigation into the use of a muscle-specific promoter to drive transgene expression was pursued. The bovine myogenin promoter and muscle creatine kinase (MCK) promoter were cloned into a lentiviral plasmid and evaluated in bovine fetal muscle cells and mouse C2C12 cells in vitro for their ability to drive GFP expression. Both promoters demonstrated an increase in GFP intensity at day nine of differentiation when compared to the nontransduced control. The retroviral basis of lentiviral plasmids has raised concern over the possible development of replication competent lentivirus (RCL). Therefore, analysis of tissues from recipients of lentivirus-treated embryos was performed to detect possible RCL. Tissues and blood serum were tested for RCL using p24 ELISA as well as qRT-PCR for the VSV-G gene. To date all tissues tested so far shown no evidence for RCL using these analyses. Analysis of offspring transgenic for an shRNA targeting myostatin will allow confirmation of RNAi as a useful tool for manipulating gene expression in large animal species. v DEDICATION This dissertation is dedicated to my family, without whom I would never have been able to make it to where I am today. My parents unending love and support have been vital throughout my educational career. I will be forever grateful for their encouragement during difficult times. My grandparents have contributed to my growth as a student and as a person and will always be in my heart. Thank you to all who have been the backbone of my growth as a student and as a person. vi ACKNOWLEDGEMENTS I would like to thank my committee chair, Dr. Westhusin, and my committee members, Dr. Long, Dr. Spencer, and Dr. Ing, for their guidance and assistance throughout the course of my career here at Texas A&M. I would also like to thank all of the present and past members of the Reproductive Sciences Lab at Texas A&M University, who were invaluable in this work. In addition, thanks to all of my family and friends for their support during this research program. vii TABLE OF CONTENTS Page ABSTRACT .............................................................................................................. iii DEDICATION .......................................................................................................... v ACKNOWLEDGEMENTS ...................................................................................... vi TABLE OF CONTENTS .......................................................................................... vii LIST OF FIGURES................................................................................................... ix LIST OF TABLES .................................................................................................... x CHAPTER I INTRODUCTION AND LITERATURE REVIEW............................ 1 Introduction .................................................................................... 1 RNA Interference ........................................................................... 2 Lentiviral Delivery of shRNAs ...................................................... 5 Off-target Effects of RNAi............................................................. 9 Large Animal Research Models..................................................... 12 Development of Transgenic Livestock .......................................... 15 Myostatin........................................................................................ 18 II IN VITRO SUPPRESSION OF MYOSTATIN.................................... 23 Introduction .................................................................................... 23 Materials and Methods ................................................................... 24 Results ............................................................................................ 34 Discussion ..................................................................................... 44 Conclusions .................................................................................... 48 III ANALYSIS OF MUSCLE-SPECIFIC PROMOTERS FOR LENTIVIRAL-MEDIATED shRNA EXPRESSION.......................... 50 Introduction .................................................................................... 50 Materials and Methods ................................................................... 51 Results ............................................................................................ 54 viii CHAPTER Page Discussion ...................................................................................... 59 Conclusions .................................................................................... 61 IV PRODUCTION OF TRANSGENIC ANIMALS WITH REDUCED MYOSTATIN EXPRESSION............................................................. 63 Introduction .................................................................................... 63 Materials and Methods ................................................................... 65 Results ........................................................................................... 68 Discussion ...................................................................................... 74 Conclusions ................................................................................... 78 V REPLICATION COMPETENT LENTIVIRUS (RCL) ANALYSIS IN RECIPIENT ANIMALS OF TRANSGENIC EMBRYOS PRODUCED BY LENTIVIRAL TRANSFER..................................... 80 Introduction .................................................................................... 80 Materials and Methods ................................................................... 82 Results ........................................................................................... 85 Discussion ..................................................................................... 90 Conclusions ................................................................................... 92 VI SUMMARY.......................................................................................... 93 REFERENCES.......................................................................................................... 96 APPENDIX A .......................................................................................................... 113 APPENDIX B .......................................................................................................... 115 VITA ......................................................................................................................... 116 ix LIST OF FIGURES FIGURE Page 1 HEK 293T cells expressing caprine myostatin as well as an shRNA targeting myostatin..................................................................................... 35 2 Suppression of myostatin mRNA in vitro.................................................. 36 3 Bovine primary muscle cells in culture...................................................... 38 4 Immunocytochemistry on bovine primary muscle cells............................. 39 5 Transduction of primary bovine fetal muscle cells .................................... 41 6 Myotube development in adult bovine
Load more

Recommended publications
  • Observation and a Numerical Study of Gravity Waves During Tropical Cyclone Ivan (2008)
    Open Access Atmos. Chem. Phys., 14, 641–658, 2014 Atmospheric www.atmos-chem-phys.net/14/641/2014/ doi:10.5194/acp-14-641-2014 Chemistry © Author(s) 2014. CC Attribution 3.0 License. and Physics Observation and a numerical study of gravity waves during tropical cyclone Ivan (2008) F. Chane Ming1, C. Ibrahim1, C. Barthe1, S. Jolivet2, P. Keckhut3, Y.-A. Liou4, and Y. Kuleshov5,6 1Université de la Réunion, Laboratoire de l’Atmosphère et des Cyclones, UMR8105, CNRS-Météo France-Université, La Réunion, France 2Singapore Delft Water Alliance, National University of Singapore, Singapore, Singapore 3Laboratoire Atmosphères, Milieux, Observations Spatiales, UMR8190, Institut Pierre-Simon Laplace, Université Versailles-Saint Quentin, Guyancourt, France 4Center for Space and Remote Sensing Research, National Central University, Chung-Li 3200, Taiwan 5National Climate Centre, Bureau of Meteorology, Melbourne, Australia 6School of Mathematical and Geospatial Sciences, Royal Melbourne Institute of Technology (RMIT) University, Melbourne, Australia Correspondence to: F. Chane Ming ([email protected]) Received: 3 December 2012 – Published in Atmos. Chem. Phys. Discuss.: 24 April 2013 Revised: 21 November 2013 – Accepted: 2 December 2013 – Published: 22 January 2014 Abstract. Gravity waves (GWs) with horizontal wavelengths ber 1 vortex Rossby wave is suggested as a source of domi- of 32–2000 km are investigated during tropical cyclone (TC) nant inertia GW with horizontal wavelengths of 400–800 km, Ivan (2008) in the southwest Indian Ocean in the upper tropo- while shorter scale modes (100–200 km) located at northeast sphere (UT) and the lower stratosphere (LS) using observa- and southeast of the TC could be attributed to strong local- tional data sets, radiosonde and GPS radio occultation data, ized convection in spiral bands resulting from wave number 2 ECMWF analyses and simulations of the French numerical vortex Rossby waves.
    [Show full text]
  • Dolly the Sheep – the First Cloned Adult Animal
    DOLLY THE SHEEP – THE FIRST CLONED ADULT ANIMAL NEW TECHNOLOGY FOR IMPROVING LIVESTOCK From Squidonius via Wikimedia Commons In 1996, University of Edinburgh scientists celebrated the birth of Dolly the Sheep, the first mammal to be cloned using SCNT cloning is the only technology adult somatic cells. The Edinburgh team’s success followed available that enables generation of 99.8% its improvements to the single cell nuclear transfer (SCNT) genetically identical offspring from selected technique used in the cloning process. individuals of adult animals (including sterilized animals). As such, it is being Dolly became a scientific icon recognised worldwide and exploited as an efficient multiplication tool SCNT technology has spread around the world and has been to support specific breeding strategies of used to clone multiple farm animals. farm animals with exceptionally high genetic The cloning of livestock enables growing large quantities of value. the most productive, disease resistant animals, thus providing more food and other animal products. Sir Ian Wilmut (Inaugural Director of MRC Centre for Regeneration and Professor at CMVM, UoE) and colleagues worked on methods to create genetically improved livestock by manipulation of stem cells using nuclear transfer. Their research optimised interactions between the donor nucleus and the recipient cytoplasm at the time of fusion and during the first cell cycle. Nuclear donor cells were held in mitosis before being released and used as they were expected to be passing through G1 phase. CLONING IN COMMERCE, CONSERVATION OF AGRICULTURE AND PRESERVATION ANIMAL BREEDS OF LIVESTOCK DIVERSITY Cloning has been used to conserve several animal breeds in the recent past.
    [Show full text]
  • Laser Interferometer Space Antenna What Is LISA?
    Volume 11 number 4 2003 FALL Quarter A FLIGHT PROGRAMS AND PROJECTS DIRECTORATE QUARTERLY PUBLICATION A Newsletter Published for Code 400 Employees INSIDE Laser Interferometer Space Antenna THIS What is LISA? ISSUE: How did the Universe begin? Does time have a beginning and an end? Does What is LISA? Page 1 space have edges? These are the questions we've struggled to answer for cen- Solar Dynamic Observatory Page 1 turies. Science and technology have now reached the point where answers to these questions are finally within our grasp. The Laser Interferometer Space Message From The Director Of Page 2 Antenna (LISA) may supply some of these answers as the mission studies Peer Award Ceremony & Picnic Page 2 the mergers of supermassive black holes, tests Einstein's Theory of General Tintypes Page 3 Relativity, probes the early Universe, and searches for gravitational waves—— its primary objective. Feedback Page 3 As the first dedicated space-based gravitational wave observatory, LISA will Technology Corner Page 6 detect waves generated by binaries within our Galaxy (the Milky Way) and Quotes of the Quarter Page 7 by massive black holes in distant galaxies. LISA will use an advanced system of laser interferometry for directly detecting and measuring them. This OBPR Free-Flyer Page 10 Best of the Best Page 12 (LISA Continued on page 4) The English Language Page 13 Peer Awards Page 14 Living with a Star Program – NASA Honor Awards Page 16 Solar Dynamics Observatory TCP Social News Page 19 Developing an understanding of the Sun took on a renewed sense of ur- Cultural Tidbits Page 19 gency during late October/early November.
    [Show full text]
  • Guide to Biotechnology 2008
    guide to biotechnology 2008 research & development health bioethics innovate industrial & environmental food & agriculture biodefense Biotechnology Industry Organization 1201 Maryland Avenue, SW imagine Suite 900 Washington, DC 20024 intellectual property 202.962.9200 (phone) 202.488.6301 (fax) bio.org inform bio.org The Guide to Biotechnology is compiled by the Biotechnology Industry Organization (BIO) Editors Roxanna Guilford-Blake Debbie Strickland Contributors BIO Staff table of Contents Biotechnology: A Collection of Technologies 1 Regenerative Medicine ................................................. 36 What Is Biotechnology? .................................................. 1 Vaccines ....................................................................... 37 Cells and Biological Molecules ........................................ 1 Plant-Made Pharmaceuticals ........................................ 37 Therapeutic Development Overview .............................. 38 Biotechnology Industry Facts 2 Market Capitalization, 1994–2006 .................................. 3 Agricultural Production Applications 41 U.S. Biotech Industry Statistics: 1995–2006 ................... 3 Crop Biotechnology ...................................................... 41 U.S. Public Companies by Region, 2006 ........................ 4 Forest Biotechnology .................................................... 44 Total Financing, 1998–2007 (in billions of U.S. dollars) .... 4 Animal Biotechnology ................................................... 45 Biotech
    [Show full text]
  • Evolution of Multiscale Vortices in the Development of Hurricane Dolly (2008)
    JANUARY 2011 F A N G A N D Z H A N G 103 Evolution of Multiscale Vortices in the Development of Hurricane Dolly (2008) JUAN FANG Key Laboratory of Mesoscale Severe Weather (MOE), Department of Atmospheric Sciences, Nanjing University, Nanjing, China FUQING ZHANG Department of Meteorology, The Pennsylvania State University, University Park, Pennsylvania (Manuscript received 6 April 2010, in final form 6 July 2010) ABSTRACT As a follow-up to a previously published article on the initial development and genesis of Hurricane Dolly (2008), this study further examines the evolution of, and interactions among, multiscale vortices ranging from the system-scale main vortex (L . 150 km) to the intermediate-scale cloud clusters (50 km , L , 150 km) and individual vorticity-rich convective cells (L , 50 km). It is found that there are apparent self-similarities among these vortices at different scales, each of which may undergo several cycles of alternating accumulation and release of convective available potential energy. Enhanced surface fluxes below individual cyclonic vortices at each scale contribute to the sustainment and reinvigoration of moist convection that in turn contributes to the maintenance and upscale growth of these vortices. Spectral analysis of horizontal divergence and relative vorticity further suggests that the cloud-cluster-scale and system-scale vortices are predominantly balanced while the individual convective vortices are largely unbalanced. The vorticity and energy produced by these individual vorticity-rich convective cells first saturate at convective scales that are subsequently transferred to larger scales. The sum of the diabatic heating released from these convective cells may be regarded as a persistent forcing on the quasi-balanced system-scale vortex.
    [Show full text]
  • The Rotary Club of Richmond
    TABLE OF CONTENTS RI President Kalyan Banerjee Bio ..................................4 INTERNATIONAL SERVICE RI President Kalyan Banerjee Letter ..............................5 2008 Refilwe Project .....................................................50 Letter from PM Stephen Harper .....................................6 Disaster Relief ...............................................................51 Letter from MP Alice Wong ...........................................7 2007 Joint Wheelchair Project .....................................52 Letter from Premier Christy Clark ..................................8 Aid for Children in Trinidad .........................................52 Letter from MLA Linda Reid ..........................................9 Korle-Bu Neuroscience .................................................53 Letter from Mayor Malcolm Brodie ..............................10 2009 Shoes for Sri Lanka ..............................................53 Letter from District Governor Hans Doge ...................11 The Rotary Foundation.................................................54 Rotary Club President Ken Whitney Bio ......................12 PolioPlus ........................................................................54 About Rotary & Paul P. Harris ......................................13 Paul Harris Fellows ........................................................55 Guiding Principles .........................................................14 Ambulance Projects ......................................................56
    [Show full text]
  • Cloning After Dolly
    Cloning after Dolly Professor Derek Burke 'Cloning' is a loose term used to describe everything from a new super race of identical near- robots (remember 'The Boys from Brazil'?) to propagating your roses. It actually means a twig, so it starts from the horticultural end, but is used much more widely. Dolly was the first cloned mammal and must be the most famous sheep in the world. Two things made her famous: first that she is an artificially induced clone, genetically identical to a previously existing sheep, for Dolly can be thought of as a vertical twin with identical genes, in this case to the parent. Second is the way in which it was done. The scientists took the nucleus from a cell from the udder of an adult sheep and implanted it in an egg cell whose original nucleus had been removed. Very surprisingly, the adult cell DNA was able to give rise to a complete individual, not just more udder cells. This was surprising because, although at the earliest stage of development the DNA is still pluripotent, capable of generating the growth of the whole animal, later in development, specialization sets in, and cells become differentiated into heart cells, liver cells etc. The whole of the genetic material is still there, but most of it is switched off and only the genes relevant to that particular cell are active. It had been thought that this process of differentiation in mammals was irreversible, so that an udder cell could only give rise to more udder cells. But we now know that differentiation can be reversed.
    [Show full text]
  • Cessation of Anadromy in a Long-Lived Fish
    Ecology, 96(7), 2015, pp. 1899–1910 Ó 2015 by the Ecological Society of America Beyond dichotomous life histories in partially migrating populations: cessation of anadromy in a long-lived fish 1,3 2 1 MORGAN H. BOND, JESSICA A. MILLER, AND THOMAS P. QUINN 1School of Aquatic and Fishery Sciences, University of Washington, Box 355020, Seattle, Washington 98195 USA 2Coastal Oregon Marine Experiment Station, Hatfield Marine Science Center, Department of Fisheries and Wildlife, Oregon State University, 2030 SE Marine Science Drive, Newport, Oregon 97365 USA Abstract. Across animal taxa, migration allows individuals to exploit habitats and resources that predictably vary seasonally in suitability. Theory predicts that the ‘‘decision’’ to migrate or not is shaped by the relative fitness costs and benefits of exhibiting a given life history. Adoption of a migratory strategy is widely thought to reflect a dichotomous outcome; individuals are either resident or migratory, and continue to exhibit this life history until death. In fishes, anadromy and freshwater residency represents a well-studied life history dichotomy. Resident individuals may adopt a migratory life history later in life, but migratory individuals are not known to abandon this pattern. Here, we investigated the fitness benefits, as measured by body size, of residency and anadromy in a salmonid fish, Dolly Varden, Salvelinus malma, in Alaska, and reveal a novel life history: cessation of migration by older, larger individuals. Otolith microchemical analysis of Dolly Varden showed that while most fish migrated to sea at least once in their lives, lifelong resident fish exist in streams with close proximity to the ocean.
    [Show full text]
  • Genetic Cloning and Testing
    Genetic Cloning and Testing BACKGROUND Genetic Cloning or years, humans successfully have used myriad genetic techniques—from GLOSSARY Fselective breeding to genetic manipulation—to create plants and animals Chromosome with desirable characteristics. Corn has been genetically engineered to improve Units in the nucleus of a cell, resistance to fungus and viral infections, rot, and drought, and calves have been composed of DNA, that contain genes; normal human somatic cells genetically altered to accelerate the growth process, to cite two examples. contain 46 chromosomes (23 pair). Clone Genetic cloning is a comparatively new process that uses the DNA contained in Duplicate. cells to “clone,” or duplicate, strands of DNA, cells, or organisms to achieve DNA Deoxyribonucleic acid; the material exact copies of the original. These genetic clones are analogous to identical inside the nucleus of a cell that human twins—two genetically identical individuals whose development can be carries genetic information. traced to the division of a single embryo. For some time, scientists have known Embryo The prefetal product of conception that multiple animals could be created in the same way that human twins are from implantation through the 12th created: by dividing a single embryo into multiple embryos shortly after fertiliza- week of development. An embryo conceived through sexual tion. This creates offspring that are genetically identical to each other and have reproduction receives half its genetic received their genetic information from each sexual parent (half from the male, information from each sexual parent—that is, half comes from half from the female). the sperm and half from the egg.
    [Show full text]
  • Cloning a Plasmid
    Dolly, herself http://oracleformsinfo.files.wordpress.com/2012/03/clone.jpg Dolly the sheep, who became famous as the first mammal to be cloned from an adult cell, has died. The news was confirmed on Friday by the Roslin Institute, the Scottish research centre which created her. A decision was taken to "euthanize" six-year-old Dolly after a veterinary examination showed that she had a progressive lung disease, the institute said in a statement. Dolly became the first mammal clone when she was born on 5 July 1996. She was revealed to the public the following year. http://martinfrost.ws/htmlfiles/roslin_dolly.html What is a clone? A clone is a • Pieces of DNA group of • Plasmids genetically • Bacteria identical … ummm….. • Eukaryotic cells in vivo • Eukaryotic cells Thingies? • Organisms • Other examples? http://media.moddb.com/images/downloads/1/25/24418/Star_wars_the_clone_wars.jpg To DNA (& etc) http://anglelakesc.blogspot.com/2011/10/road-construction-detours.html Cloning a plasmid http://wiki.biomine.skelleftea.se/wiki/images/9/93/Cloning_with_the_help_of_LacZ_.png Do clones occur in nature? • Identical twins • Lymphocytes • An embryo … http://cadencemag.com/wp-content/uploads/2012/03/twins.gif YOU are a clone of your original fertilized egg! Are cloned cells identical? -- genetically? -- functionally? How? Why? Why not? Dolly & her baby – so Dolly was fertile Fertility is a common measure of whether a genome is really complete http://news.bbc.co.uk/2/hi/science/nature/2764039.stm First Cloned Mouse Dies Of Old Age: Technique Believed More Reliable Than That Used For Dolly The Sheephttp://www.cbsnews.com/stories/2000/05/10/tech/main193799.shtml • Why clone? • Clone what ? • Humans – Whole? • Financial benefit – Parts? • Convenience • Animals • Uniformity – Agricultural • Personal emotion – Environmental restoration • Medical intervention – Fun – e.g.
    [Show full text]
  • BSCI410-Liu/SP09 Midterm Exam #2/Apr. 14 Your Name
    BSCI410-Liu/SP09 Midterm Exam #2/Apr. 14 Your Name: 1. (28 points total) Scientists are trying to unlock the molecular basis of a mysterious hereditary disease named DEAN SYMDROM (DS). ds mutant mice show a lack of memory and ds/ds mutant mice, when crossed with wild type (+/+) mice, produces progeny that also show memory loss. (a) (4 points) Is the ds mutation recessive or dominant? dominant (b) (4 points) Using molecular markers such as SNPs, scientists have mapped the putative DS gene to a small chromosomal region. This region contains three genes: X, Y, and Z. Design an experiment to test the expression of X, Y, and Z genes. Which tissue do you expect the candidate DS gene to be expressed in? Northern blot, RT-PCR, in situ hybridization to see which gene is expressed in brain tissue (c) (8 points) Through the above experiments, scientists identified gene Y that is expressed in the expected tissue. List two additional experiments that will definitely prove that gene Y is the DS gene. 1) sequence mutant mice DNA and compare with WT. If one detects a mutation in Y in the mutant mice, it would prove it. 2) Transgenic study - Introduce the mutant gene into the WT mouse to see if it causes memory loss (cannot introduce WT into mutant mouse to rescue because the mutation is dominant) I also took other reasonable answers. NOTE: Testing RNA expression will not definitely prove that gene Y is the DS gene. (d) (8 points) After identifying the mouse DS gene, you now have the DNA sequence of the mouse DS gene.
    [Show full text]
  • Gender Equality, HIV, and AIDS: Challenges for the Education Sector
    Gender Equality and HIV and AIDS A Challenge for the Education Sector Edited by Sheila Aikman, Elaine Unterhalter and Tania Boler Gender Equality, HIV, and AIDS A Challenge for the Education Sector Oxfam GB Oxfam GB, founded in 1942, is a development, humanitarian, and campaigning agency dedicated to finding lasting solutions to poverty and suffering around the world. Oxfam believes that every human being is entitled to a life of dignity and opportunity, and it works with others worldwide to make this become a reality. From its base in Oxford in the United Kingdom, Oxfam GB publishes and distributes a wide range of resource materials for development and relief workers, researchers and campaigners, schools and colleges, and the general public, as part of its programme of advocacy, education, and communications. Oxfam GB is a member of Oxfam International, a confederation of 13 agencies of diverse cultures and languages, which share a commitment to working for an end to injustice and poverty – both in long-term development work and at times of crisis. For further information about Oxfam’s publishing, and online ordering, visit www.oxfam.org.uk/publications For information about Oxfam’s development, advocacy, and humanitarian relief work around the world, visit www.oxfam.org.uk Gender Equality, HIV, and AIDS A Challenge for the Education Sector Edited by Sheila Aikman, Elaine Unterhalter, and Tania Boler Front cover: Children at play on their way home from school in Kitwe, Zambia. (Annie Bungeroth/Oxfam) The views expressed are those of the authors and not necessarily those of the publishers. First published by Oxfam GB and ActionAid in 2008 © Oxfam GB 2008 ISBN 978-085598-586-8 A catalogue record for this publication is available from the British Library.
    [Show full text]