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Gene Use Restriction Technologies for Transgenic Plant Bioconfinement
Plant Biotechnology Journal (2013) 11, pp. 649–658 doi: 10.1111/pbi.12084 Review article Gene use restriction technologies for transgenic plant bioconfinement Yi Sang, Reginald J. Millwood and C. Neal Stewart Jr* Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA Received 1 February 2013; Summary revised 3 April 2013; The advances of modern plant technologies, especially genetically modified crops, are considered accepted 9 April 2013. to be a substantial benefit to agriculture and society. However, so-called transgene escape *Correspondence (fax 1-865-974-6487; remains and is of environmental and regulatory concern. Genetic use restriction technologies email [email protected]) (GURTs) provide a possible solution to prevent transgene dispersal. Although GURTs were originally developed as a way for intellectual property protection (IPP), we believe their maximum benefit could be in the prevention of gene flow, that is, bioconfinement. This review describes the underlying signal transduction and components necessary to implement any GURT system. Keywords: transgenic plants, Furthermore, we review the similarities and differences between IPP- and bioconfinement- transgene escape, male sterility, oriented GURTs, discuss the GURTs’ design for impeding transgene escape and summarize embryo sterility, transgene deletion, recent advances. Lastly, we go beyond the state of the science to speculate on regulatory and gene flow. ecological effects of implementing GURTs for bioconfinement. Introduction proposed (Daniell, 2002; Gressel, 1999; Moon et al., 2011), including strategies for male sterility (Mariani et al., 1990), Transgenic crops have become an integral part of modern maternal inheritance (Daniell et al., 1998; Iamtham and Day, agriculture and have been increasingly adopted worldwide (James, 2000; Ruf et al., 2001), transgenic mitigation (Al-Ahmad et al., 2011). -
Gene Therapy and Genetic Engineering: Frankenstein Is Still a Myth, but It Should Be Reread Periodically
Indiana Law Journal Volume 48 Issue 4 Article 2 Summer 1973 Gene Therapy and Genetic Engineering: Frankenstein is Still a Myth, but it Should be Reread Periodically George A. Hudock Indiana University - Bloomington Follow this and additional works at: https://www.repository.law.indiana.edu/ilj Part of the Genetics and Genomics Commons Recommended Citation Hudock, George A. (1973) "Gene Therapy and Genetic Engineering: Frankenstein is Still a Myth, but it Should be Reread Periodically," Indiana Law Journal: Vol. 48 : Iss. 4 , Article 2. Available at: https://www.repository.law.indiana.edu/ilj/vol48/iss4/2 This Article is brought to you for free and open access by the Law School Journals at Digital Repository @ Maurer Law. It has been accepted for inclusion in Indiana Law Journal by an authorized editor of Digital Repository @ Maurer Law. For more information, please contact [email protected]. GENE THERAPY AND GENETIC ENGINEERING: FRANKENSTEIN IS STILL A MYTH, BUT IT SHOULD BE REREAD PERIODICALLY GEORGE A. HUDOCKt Biotechnology and the law are far removed from each other as disciplines of human intellect. Yet the law and my own discipline, genetics, have come together in many courtrooms concerning such matters as paternity, and they will continue to intersect with increasing frequency as the visions of 100 years ago become the reality of today. This article examines the implications of recent research for human genetic therapy and genetic engineering, and suggests some guidelines for legal regulation of genetic technology. The following discussion derives from three premises which I view as basic: (1) that which is currently possible in genetic engineering, and in fact has already been done, is generally underestimated; (2) what may be possible in the near future is quite commonly overesti- mated; (3) regulation of the application of genetic technology is possible and will not be overwhelmingly complicated. -
Gibson Assembly Cloning Guide, Second Edition
Gibson Assembly® CLONING GUIDE 2ND EDITION RESTRICTION DIGESTFREE, SEAMLESS CLONING Applications, tools, and protocols for the Gibson Assembly® method: • Single Insert • Multiple Inserts • Site-Directed Mutagenesis #DNAMYWAY sgidna.com/gibson-assembly Foreword Contents Foreword The Gibson Assembly method has been an integral part of our work at Synthetic Genomics, Inc. and the J. Craig Venter Institute (JCVI) for nearly a decade, enabling us to synthesize a complete bacterial genome in 2008, create the first synthetic cell in 2010, and generate a minimal bacterial genome in 2016. These studies form the framework for basic research in understanding the fundamental principles of cellular function and the precise function of essential genes. Additionally, synthetic cells can potentially be harnessed for commercial applications which could offer great benefits to society through the renewable and sustainable production of therapeutics, biofuels, and biobased textiles. In 2004, JCVI had embarked on a quest to synthesize genome-sized DNA and needed to develop the tools to make this possible. When I first learned that JCVI was attempting to create a synthetic cell, I truly understood the significance and reached out to Hamilton (Ham) Smith, who leads the Synthetic Biology Group at JCVI. I joined Ham’s team as a postdoctoral fellow and the development of Gibson Assembly began as I started investigating methods that would allow overlapping DNA fragments to be assembled toward the goal of generating genome- sized DNA. Over time, we had multiple methods in place for assembling DNA molecules by in vitro recombination, including the method that would later come to be known as Gibson Assembly. -
Cloning of Gene Coding Glyceraldehyde-3-Phosphate Dehydrogenase Using Puc18 Vector
Available online a t www.pelagiaresearchlibrary.com Pelagia Research Library European Journal of Experimental Biology, 2015, 5(3):52-57 ISSN: 2248 –9215 CODEN (USA): EJEBAU Cloning of gene coding glyceraldehyde-3-phosphate dehydrogenase using puc18 vector Manoj Kumar Dooda, Akhilesh Kushwaha *, Aquib Hasan and Manish Kushwaha Institute of Transgene Life Sciences, Lucknow (U.P), India _____________________________________________________________________________________________ ABSTRACT The term recombinant DNA technology, DNA cloning, molecular cloning, or gene cloning all refers to the same process. Gene cloning is a set of experimental methods in molecular biology and useful in many areas of research and for biomedical applications. It is the production of exact copies (clones) of a particular gene or DNA sequence using genetic engineering techniques. cDNA is synthesized by using template RNA isolated from blood sample (human). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) is one of the most commonly used housekeeping genes used in comparisons of gene expression data. Amplify the gene (GAPDH) using primer forward and reverse with the sequence of 5’-TGATGACATCAAGAAGGTGGTGAA-3’ and 5’-TCCTTGGAGGCCATGTGGGCCAT- 3’.pUC18 high copy cloning vector for replication in E. coli, suitable for “blue-white screening” technique and cleaved with the help of SmaI restriction enzyme. Modern cloning vectors include selectable markers (most frequently antibiotic resistant marker) that allow only cells in which the vector but necessarily the insert has been transfected to grow. Additionally the cloning vectors may contain color selection markers which provide blue/white screening (i.e. alpha complementation) on X- Gal and IPTG containing medium. Keywords: RNA isolation; TRIzol method; Gene cloning; Blue/white screening; Agarose gel electrophoresis. -
New Techniques of Genetic Engineering
March 2016 New techniques of genetic engineering Why EU GMO law must be fully applied to the so-called ‘New Plant Breeding Techniques’ The European Commission is considering whether genetically modified organisms (GMOs) that have been produced through a range of new techniques should be excluded from the European Union’s GMO regulations. Biotechnology companies want to apply these techniques to engineer plants and animals for use in industrial food, biomass and biofuel production. They argue that these new methods to directly modify the genetic make-up of living organisms fall outside the scope of EU GMO regulations. This would mean that there is no risk assessment, labelling and monitoring of GM organisms produced by the new techniques and their derived products. The Commission has announced that it will present a legal analysis on the matter by the end of March 2016. The new GMOs present a real risk to the environment and human health. Legal analysis shows that they are covered by EU GMO law. If they were to escape EU regulations, any potential negative effects on food, feed or environmental safety would go unchecked. European consumers, farmers and breeders would have no way to avoid GMOs. The Commission should leave no doubt that all products of genetic engineering are subject to EU GMO law which requires rigorous risk assessment, detectability and labelling. 1 Which techniques are we talking about? The biotechnology industry and the European Commission use the term ‘New Plant Breeding Techniques’ to refer to a diverse set of genetic -
Drawing the Line: Disability, Genetic Intervention and Bioethics
laws Article Drawing the Line: Disability, Genetic Intervention and Bioethics Adam Conti Graduate student, Melbourne Law School, University of Melbourne, 185 Pelham St., Carlton, VIC 3053, Australia; [email protected] Received: 2 June 2017; Accepted: 10 July 2017; Published: 17 July 2017 Abstract: Meteoric scientific advances in genetic technologies with the potential for human gene editing intervention pose tremendous legal, medical, social, ethical and moral issues for society as a whole. Persons with disabilities in particular have a significant stake in determining how these technologies are governed at the international, domestic and individual levels in the future. However, the law cannot easily keep up with the rate of scientific progression. This paper aims to posit a methodology of reform, based on a core value of human dignity, as the optimal course of action to ensure that the interests of persons with disabilities, other possibly marginalised groups, and the scientific community, are balanced fairly. The paper critically analyses the current law and varying bioethical perspectives to ultimately conclude that a clear principled approach toward open discussion and consensus is of paramount importance to have any chance of devising an effective regulatory regime over human gene editing technology. Keywords: disability; human rights; genetics; gene editing; bioethics; governance; human dignity; eugenics; germline; Convention on the Rights of Persons with Disabilities The true good is in the different, not the same (Menand 2004). 1. Introduction Popular, professional and scholarly interest in genetics and their influence on human variability, behaviour and development has grown exponentially in recent years. In no small part has this interest been bolstered by mainstream media coverage of large-scale collaborative scientific initiatives like the Human Genome Project, which endeavoured to identify and map the human genome and determine the sequence of nucleotide base pairs that make up our DNA. -
The Transgenic Core Facility
Genetic modification of the mouse Ben Davies Wellcome Trust Centre for Human Genetics Genetically modified mouse models • The genome projects have provided us only with a catalogue of genes - little is know regarding gene function • Associations between disease and genes and their variants are being found yet the biological significance of the association is frequently unclear • Genetically modified mouse models provides a powerful method of assaying gene function in the whole organism Sequence Information Human Mutation Gene of Interest Mouse Model Reverse Genetics Phenotype Gene Function How to study gene function Human patients: • Patients with gene mutations can help us understand gene function • Human’s don’t make particularly willing experimental organisms • An observational science and not an experimental one • Genetic make-up of humans is highly variable • Difficult to pin-point the gene responsible for the disease in the first place Mouse patients: • Full ability to manipulate the genome experimentally • Easy to maintain in the laboratory – breeding cycle is approximately 2 months • Mouse and human genomes are similar in size, structure and gene complement • Most human genes have murine counterparts • Mutations that cause disease in human gene, generally produce comparable phentoypes when mutated in mouse • Mice have genes that are not represented in other model organisms e.g. C. elegans, Drosophila – genes of the immune system What can I do with my gene of interest • Gain of function – Overexpression of a gene of interest “Transgenic -
GFP Plasmid) 35X
Název: Biotechnology and Fluorescent protein Školitel: Ana Maria Jimenez Jimenez, Iva Blažková Datum: 19.7.2013 Reg.č.projektu: CZ.1.07/2.3.00/20.0148 Název projektu: Mezinárodní spolupráce v oblasti "in vivo" zobrazovacích technik INTRODUCTION GFP - Green fluorescent protein A protein composed of 238 amino acid residues (26.9 kDa) exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range GFP was isolated from the jellyfish Aequorea victoria. In cell and molecular biology, the GFP gene is frequently used as a reporter of expression The GFP gene has been introduced and expressed in many bacteria, yeast and other fungi, fish, plant and mammalian cells. Green fluorescent protein (GFP) Nobel prize in Chemistry (2008): for the discovery and development of the green fluorescent protein, GFP Osamu Shimomura Martin Chalfie Roger Y. Tsien Now GFP is found in laboratories all over the world where it is used in every conceivable plant and animal GFP Types Fluorescent proteins enable the creation of highly specific biosensors to monitor a wide range of intracellular phenomena. Mutagenesis of A. victoria GFP has resulted in fluorescent proteins that range in color from blue to yellow Transluminator Fluoresence spectrophotometry Fluorescence In-vivo Xtreme Detection Fluorescence microscopy In-vivo Xtreme cm Polyacrylamide gel In-vivo Xtreme 1/16 1/8 1/4 1/2 1 10000 y = 9003x + 732,46 9000 R² = 0,9984 Max-Backgroumd 8000 7000 Rostoucí koncentrace Mean-Background 6000 5000 4000 3000 y = 5748,2x + 490,64 R² = 0,9995 2000 Fluorescence intensity [a.u.] 1000 0 0 0,2 0,4 0,6 0,8 1 1,2 concentration [µg/ml] In-vivo Xtreme GFP encapsulated GFP in liposome GFP water in liposome Max.int: 8800 a. -
Review on Applications of Genetic Engineering and Cloning in Farm Animals
Journal of Dairy & Veterinary Sciences ISSN: 2573-2196 Review Article Dairy and Vet Sci J Volume 4 Issue 1 - October 2017 Copyright © All rights are reserved by Ayalew Negash DOI: 10.19080/JDVS.2017.04.555629 Review on Applications of Genetic Engineering And Cloning in Farm animals Eyachew Ayana1, Gizachew Fentahun2, Ayalew Negash3*, Fentahun Mitku1, Mebrie Zemene3 and Fikre Zeru4 1Candidate of Veterinary medicine, University of Gondar, Ethiopia 2Candidate of Veterinary medicine, Samara University, Ethiopia 3Lecturer at University of Gondar, University of Gondar, Ethiopia 4Samara University, Ethiopia Submission: July 10, 2017; Published: October 02, 2017 *Corresponding author: Ayalew Negash, Lecturer at University of Gondar, College of Veterinary Medicine and science, University of Gondar, P.O. 196, Gondar, Ethiopia, Email: Abstract Genetic engineering involves producing transgenic animal’s models by using different techniques such as exogenous pronuclear DNA highly applicable and crucial technology which involves increasing animal production and productivity, increases animal disease resistance andmicroinjection biomedical in application. zygotes, injection Cloning ofinvolves genetically the production modified embryonicof animals thatstem are cells genetically into blastocysts identical and to theretrovirus donor nucleus.mediated The gene most transfer. commonly It is applied and recent technique is somatic cell nuclear transfer in which the nucleus from body cell is transferred to an egg cell to create an embryo that is virtually identical to the donor nucleus. There are different applications of cloning which includes: rapid multiplication of desired livestock, and post-natal viabilities. Beside to this Food safety, animal welfare, public and social acceptance and religious institutions are the most common animal conservation and research model. -
Molecular Biology and Applied Genetics
MOLECULAR BIOLOGY AND APPLIED GENETICS FOR Medical Laboratory Technology Students Upgraded Lecture Note Series Mohammed Awole Adem Jimma University MOLECULAR BIOLOGY AND APPLIED GENETICS For Medical Laboratory Technician Students Lecture Note Series Mohammed Awole Adem Upgraded - 2006 In collaboration with The Carter Center (EPHTI) and The Federal Democratic Republic of Ethiopia Ministry of Education and Ministry of Health Jimma University PREFACE The problem faced today in the learning and teaching of Applied Genetics and Molecular Biology for laboratory technologists in universities, colleges andhealth institutions primarily from the unavailability of textbooks that focus on the needs of Ethiopian students. This lecture note has been prepared with the primary aim of alleviating the problems encountered in the teaching of Medical Applied Genetics and Molecular Biology course and in minimizing discrepancies prevailing among the different teaching and training health institutions. It can also be used in teaching any introductory course on medical Applied Genetics and Molecular Biology and as a reference material. This lecture note is specifically designed for medical laboratory technologists, and includes only those areas of molecular cell biology and Applied Genetics relevant to degree-level understanding of modern laboratory technology. Since genetics is prerequisite course to molecular biology, the lecture note starts with Genetics i followed by Molecular Biology. It provides students with molecular background to enable them to understand and critically analyze recent advances in laboratory sciences. Finally, it contains a glossary, which summarizes important terminologies used in the text. Each chapter begins by specific learning objectives and at the end of each chapter review questions are also included. -
Schematic and Time Line for the Generation of Knockout Mice Aurora Burds Connor, Feb 2007
Schematic and Time Line for the Generation of Knockout Mice Aurora Burds Connor, Feb 2007 Making the DNA construct Time Line (in your lab) The Transgenic Facility also has a “Beginner’s Guide to Gene Targeting” on the website in Methods. We are also happy to assist with advice and reagents to help you make an effective targeting construct. It is recommended that you contact Aurora ([email protected]) early in the process. Acquire the genomic DNA for your gene or locus of interest You can either screen a 129/Sv genomic library 1-3 months or use genomic databases, public BACs and PCR for a C57BL/6 background Make a DNA construct. DNA from the genomic locus (orange) flanks the DNA to be inserted (blue) and it is placed in a bacterial plasmid. This construct is designed to add new pieces of DNA into the mouse genome at a 3-6 months desired locus. In a knockout experiment, the new DNA will replace the normal gene. Linearize the construct and give it to the Rippel ES Cell Facility at MIT Generating Targeted ES Cells Time Line (done by the Facility) Day 0 Insert the DNA into embryonic stem cells (ES cells) via electroporation. In the cells, the homologous pieces of DNA recombine. This inserts new DNA from the construct into the desired locus, usually replacing a portion of the original genome. Place the ES cells in selective media, allowing for the growth of cells containing the DNA construct. - The DNA construct has a drug-resistance marker Day 1-10 - Very few of the cells take up the construct – cells that do not will die because they are not resistant to the drug added to the media. -
Genetic Engineering & Genetically Modified Organisms
Genetic Engineering & Genetically Modified Organisms: Forming Informed Opinions By Smith, Lisa Scientific Theme(s): Science and Technology *Relationships among science, technology, and society Grade Level(s): 6-8 Lesson Duration: Designed for one 70 minute lesson Overview This lesson provides students an introduction to genetic engineering and genetically modified organisms, and raises student awareness of the potential Bias of availaBle information and the importance of forming informed, defendaBle opinions regarding controversial topics in science. The lesson was designed as an introduction for students to the genetic engineering and genetically modified organisms in preparation for a two-week research project exploring GMO topics, culminating with student presentations giving their opinions on the topics. Objectives Students will: 1. Define the term, ‘Biotechnology’. 2. Understand the difference Between selective Breeding and genetic engineering. 3. Identify different applications of genetic engineering, and recognize that all genetically engineered organisms are genetically modified organisms (GMOs). 4. Recognize Bias in print media, and to Be aware of the need to identify sources. 5. Understand the importance of forming informed, defendaBle opinions. Grade Level Expectations (GLEs) Addressed Science as Inquiry and Process [7] SA2.1 identifying and evaluating the sources used to support scientific statements Science and Technology [7] SE1.1 descriBing how puBlic policy affects the student’s life (e.g., puBlic waste disposal) [7] SE3.1 recognizing the effects of a past scientific discovery, invention, or scientific Breakthrough (e.g., DDT, internal comBustion engine) Required BacKground This lesson builds upon concepts covered in previous lessons on DNA, genes, and heredity. Students should have a solid understanding of the Basics of these concepts, including the idea that DNA is the “Blueprint” of life, that genes are coding regions of DNA, and that traits encoded By genes can Be inherited.