Gene Editing Reflections from the Panel Co-Chairs Scenario Summaries + Scenarios (Healthcare, Pest Control and Primary Industries) Legal and Regulatory Implications

Total Page:16

File Type:pdf, Size:1020Kb

Gene Editing Reflections from the Panel Co-Chairs Scenario Summaries + Scenarios (Healthcare, Pest Control and Primary Industries) Legal and Regulatory Implications ROYAL SOCIETY TE APĀRANGI GENE EDITING REFLECTIONS FROM THE PANEL CO-CHAIRS SCENARIO SUMMARIES + SCENARIOS (HEALTHCARE, PEST CONTROL AND PRIMARY INDUSTRIES) LEGAL AND REGULATORY IMPLICATIONS AUGUST 2019 TŪHURA TOROHĒ TOHATOHA CONTENTS GENE EDITING REFLECTIONS FROM THE PANEL CO-CHAIRS GENE EDITING GENE EDITING SCENARIOS IN SCENARIOS HEALTHCARE SUMMARY IN HEALTHCARE GENE EDITING GENE EDITING SCENARIOS IN PEST SCENARIOS CONTROL SUMMARY IN PEST CONTROL GENE EDITING GENE EDITING SCENARIOS IN THE SCENARIOS PRIMARY INDUSTRIES IN THE PRIMARY SUMMARY INDUSTRIES GENE EDITING LEGAL AND REGULATORY IMPLICATIONS ROYAL SOCIETY TE APĀRANGI GENE EDITING REFLECTIONS FROM THE PANEL CO-CHAIRS BARRY SCOTT AND DAVID PENMAN AUGUST 2019 Nā te iahia kia titiro, ā, ka kite ai tātou te mutunga. You must understand the beginning if you wish to see the end. 02 | ROYAL SOCIETY TE APĀRANGI GENE EDITING REFLECTIONS FROM THE PANEL CO-CHAIRS | 03 GENE EDITING: REFLECTIONS FROM THE PANEL CO-CHAIRS BARRY SCOTT AND DAVID PENMAN Random mutagenesis and selection in nature In response to these advances, many reports have has underpinned evolution and diversity of all been released and summits organised by academies life and the resulting domestication of plants and and research organisations around the world to animals. In modern times, advances in science and explain the technology, the context in which it is technology have allowed humankind to augment this being used, the issues that arise for society, and natural process in increasingly sophisticated ways the impact of these scientific advances on current through selective breeding programmes and the regulatory frameworks. Prominent among these was use of techniques such as irradiation and chemical the “International Summit on Human Gene Editing” mutagenesis to enhance the rate of gene mutation. organised by the US National Academies of Sciences and Medicine, the Royal Society (London) and the The development of a number of DNA technologies, Chinese Academy of Sciences in late 2015 to discuss and our ability to sequence entire genomes, has guidelines for the use of gene editing in humans. opened the door to modifying specific genes to generate new traits and characteristics. The These international reviews and summits have publication by Doudna, Charpentier and colleagues been immensely helpful in informing the public, in 2012, demonstrating how a bacterial system researchers and regulatory bodies around the world, for adaptive immunity called CRISPR-Cas9 could including New Zealand, and providing a framework be engineered to precisely edit genomes, has set for engaging internationally. However, New Zealand in motion a revolution in biology. It has been quite needs to have its own perspective given our unique astounding how quickly laboratories around the cultural heritage and environment, the special world have adopted this new tool for applications challenges we face in maintaining our biodiversity across biology, from modifying plants, to altering and a viable and productive primary industry, and insect development and potential treatment our unique regulatory environment. Furthermore, of some human diseases. The relative ease with there has been no review of gene technologies in which genomes can now be sequenced and edited New Zealand since the Royal Commission on Genetic has generated considerable excitement within the Modification held in 2001 and the subsequent scientific community. However, it has also raised amendments to the Hazardous Substances and New significant concerns about the social, legal and Organisms Act (1996). The field of genome science ethical issues raised by the use of the technology, has advanced dramatically since then, especially none more so than the potential to edit genes the ability to sequence organism genomes and to in human embryos. manipulate those genomes in a very precise way. GENE EDITING REFLECTIONS FROM THE PANEL CO-CHAIRS | 03 ROYAL SOCIETY TE APĀRANGI GENE EDITING PANEL AND MĀORI REFERENCE GROUP Royal Society Te Apārangi, as an independent The panel chose to consider the implications science body, has a function under its Act to of the technology in parallel work streams using provide expert advice on important public issues a range of scenarios in three areas: healthcare, to the Government and the community. In 2016, environmental pest management and primary the Society initiated a programme of work to explore industries. The scenarios are illustrative – they are the implications of gene editing technology for New not panel recommendations for priorities for New Zealand, motivated by the importance of this rapidly Zealand application. They are presented in a stepped advancing science, the need to raise awareness of approach of increasing potential risk averseness its potential applications, and to support informed and near and long-term benefits, to challenge and discussion and debate about its implications for promote public engagement, and test the current New Zealanders. regulatory regime. Each set of scenarios aimed to consider potential ethical, cultural and legal issues The first output of this programme was a short alongside the opportunities and potential risks and document entitled “Gene editing. Evidence benefits. This approach proved to be a productive update”, released in November 2016. This provided one for initiating a conversation with the New background on gene modification technologies Zealand public. and their evolution for the media, educators, policy makers and the public. Royal Society Te Apārangi Sitting behind the scenarios are technical papers then convened a multidisciplinary panel of experts, that provide a more comprehensive overview of the supported by a Māori reference group, to consider research evidence base and implications for each the social, cultural, legal and economic implications of the three areas. These are fully referenced for of gene-editing technologies for New Zealand. This readers to access the primary literature relevant to paper outlines the approach of the panel and makes each area considered, and have been peer reviewed some concluding observations. nationally and internationally. The panel was not asked to come to a view about New territory for Royal Society Te Apārangi was the merits or otherwise of any particular application to enlist the support of a Māori Reference Group of gene editing. Rather, its role has been to provide to assist the panel in capturing Māori views and information and resources that will allow others to approaches to assessing this technology. While the have well informed discussions and debates. Indeed, papers reflect particular ways in which some Māori one of the panel’s main observations is that there would assess their use of gene editing technology, is an urgent need for wide discussion and debate the panel observed wide diversity in views across about gene editing within and across all New Zealand both Māori and non-Māori communities. communities, as global research and development in applications of gene editing is continuing apace. Some countries are reviewing, or have already reviewed, their regulations in response to these developments. 04 | ROYAL SOCIETY TE APĀRANGI GENE EDITING REFLECTIONS FROM THE PANEL CO-CHAIRS | 05 PUBLIC FEEDBACK Other innovations for Royal Society Te Apārangi • In the primary industries, comments on the in this process were the publication of material benefits of using gene-editing technology in a series of discrete work pieces over time rather included that it could provide a useful tool than one large report, and initially publishing for supporting competitive advantage, and for the scenarios in draft form to allow feedback. protecting New Zealand’s flora and fauna. There This enabled the panel to undertake a series of were concerns about unintended consequences, engagement workshops around the country to seek a need for better understanding of the relevant feedback and identify additional information that genetics, and that use of gene-editing technology could be covered in the final technical papers. Senior would compromise the New Zealand brand and school students attended some of these sessions any “GM free” competitive advantage. and the panel invited informal comment via the • Across all scenarios, feedback from Māori Society’s website. Two hui were specifically aimed participants highlighted the importance of at Māori communities. whakapapa and mauri, involving tangata whenua This feedback process targeted testing the around indigenous species, protection of data, and information in the documents for completeness intellectual property implications of gene editing and usefulness – undertaking a comprehensive taonga species. national consultation was beyond the resources • Royal Society Te Apārangi was criticised on and mandate of Royal Society Te Apārangi and the occasions for appearing to take an advocacy panel. A number of themes were apparent from position on gene editing through its publication these interactions: of scenarios. • For all three themes, there were views for and • The Society also received considerable positive against the use of gene editing. feedback on undertaking the work and its use of scenarios. The society was often encouraged to • In healthcare, there was an appetite to consider lead a much wider engagement with communities certain therapeutic gene-editing applications given its independence and scientific standing. as long as it was safe enough to rule out negative side effects, and that it would enhance human • The
Recommended publications
  • CRISPR/Cas9-Mediated Genome Editing Efficiently Creates Specific
    www.nature.com/scientificreports Correction: Author Correction OPEN CRISPR/Cas9-mediated genome editing efciently creates specifc mutations at multiple loci using one Received: 18 April 2017 Accepted: 3 July 2017 sgRNA in Brassica napus Published: xx xx xxxx Hong Yang1, Jia-Jing Wu1, Ting Tang1, Ke-De Liu 2 & Cheng Dai1 CRISPR/Cas9 is a valuable tool for both basic and applied research that has been widely applied to diferent plant species. Nonetheless, a systematical assessment of the efciency of this method is not available for the allotetraploid Brassica napus—an important oilseed crop. In this study, we examined the mutation efciency of the CRISPR/Cas9 method for 12 genes and also determined the pattern, specifcity and heritability of these gene modifcations in B. napus. The average mutation frequency for a single-gene targeted sgRNA in the T0 generation is 65.3%. For paralogous genes located in conserved regions that were targeted by sgRNAs, we observed mutation frequencies that ranged from 27.6% to 96.6%. Homozygotes were readily found in T0 plants. A total of 48.2% of the gene mutations, including homozygotes, bi-alleles, and heterozygotes were stably inherited as classic Mendelian alleles in the next generation (T1) without any new mutations or reversions. Moreover, no mutation was found in the putative of-target sites among the examined T0 plants. Collectively, our results demonstrate that CRISPR/Cas9 is an efcient tool for creating targeted genome modifcations at multiple loci that are stable and inheritable in B. napus. These fndings open many doors for biotechnological applications in oilseed crops.
    [Show full text]
  • RNA-Guided CRISPR-Cas Technologies for Genome-Scale Investigation of Disease Processes Sean E Humphrey and Andrea L Kasinski*
    Humphrey and Kasinski Journal of Hematology & Oncology (2015) 8:31 DOI 10.1186/s13045-015-0127-3 JOURNAL OF HEMATOLOGY & ONCOLOGY REVIEW Open Access RNA-guided CRISPR-Cas technologies for genome-scale investigation of disease processes Sean E Humphrey and Andrea L Kasinski* Abstract From its discovery as an adaptive bacterial and archaea immune system, the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas system has quickly been developed into a powerful and groundbreaking programmable nuclease technology for the global and precise editing of the genome in cells. This system allows for comprehensive unbiased functional studies and is already advancing the field by revealing genes that have previously unknown roles in disease processes. In this review, we examine and compare recently developed CRISPR-Cas platforms for global genome editing and examine the advancements these platforms have made in guide RNA design, guide RNA/Cas9 interaction, on-target specificity, and target sequence selection. We also explore some of the exciting therapeutic potentials of the CRISPR-Cas technology as well as some of the innovative new uses of this technology beyond genome editing. Introduction from a region of the host genome adjacent to the Clustered regularly interspaced short palindromic repeats CRISPR region is directed to the invading DNA in a (CRISPR)-Cas systems are adaptive immune systems used sequence-dependent manner via the crRNA. Once by many bacteria and archaea to fight off foreign DNA in bound to the foreign DNA, Cas9 introduces a double- the form of bacterial phages and/or plasmids [1-5]. Al- stranded break in the foreign DNA [11-13].
    [Show full text]
  • CRISPR/Cas9 Genome Editing Brochure
    mirusbio.com Cas9 Target Sequence Guide RNA GENOME EDITING: CRISPR/CAS9 DELIVERY METHODS GENOME EDITING: CRISPR/CAS9 DELIVERY What is CRISPR/Cas9 Genome Editing? The CRISPR/Cas9 system is a powerful tool for genome editing in mammalian cells that allows researchers to generate genetic variants at lower cost and with higher throughput than alternative methods like zinc finger nuclease (ZFN) or transcription activator-like effector nuclease (TALEN) genome editing. Cas9 PAM Genomic DNA Target Sequence Guide RNA crRNA tracrRNA The CRISPR/Cas9 RNP Complex. The CRISPR associated protein 9 (Cas9) endonuclease (blue) is targeted to DNA by a guide RNA (gRNA), which can be supplied as a two-part system consisting of CRISPR RNA (crRNA) and trans-activating crRNA (tracrRNA) or as a single guide RNA (sgRNA), where the crRNA and tracrRNA are connected by a linker (dotted line). Target recognition is facilitated by the protospacer-adjacent motif (PAM). A double strand break (DSB) occurs 3 bp upstream of the PAM. CRISPR Facilitates Multiple Types of Genome Modification Cleavage of Target DNA By Cas9 Deletion Modication Insertion Multiple Genomic Alterations are Possible Following Cleavage of Target DNA by Cas9. Variable length insertions and/ or deletions (indels) can result near the DNA break due to mistakes in DNA repair by the endogenous non-homologous end joining (NHEJ) pathway. These indels frequently result in disruption of gene function. Alternatively, by supplying a DNA repair template, researchers can leverage the homology-directed repair (HDR) pathway to create defined deletions, insertions or other modifications. 2 TO ORDER | Toll Free 888.530.0801 | Direct 608.441.2852 | www.mirusbio.com Glossary of CRISPR Terms Term Definition CRISPR Associated Protein 9 - Cas9 is an RNA-guided DNA endonuclease from the type Cas9 II CRISPR system of Streptococcus pyogenes that has been adapted for use in genome editing applications.
    [Show full text]
  • Download Gene Editing Evidence Update
    Gene Editing Evidence Update Summary The three most widely used new gene-editing tools Gene editing involves the insertion, deletion use bacterial proteins to find, cut, edit, add or replace or replacement of genetic material called DNA. genes, and are known as Zinc Fingers (ZFNs), TALENs, New gene-editing technologies have been developed and CRISPR. which have increased the speed, ease and accuracy Gene-editing technologies open up new opportunities of making changes to DNA in cells, and their use is and potential risks from new uses which may challenge increasing rapidly. people’s views on what is acceptable. These technologies are beginning to be used for new These new technologies pose challenges for regulators approaches in a variety of areas including research, who will find it harder to distinguish between genetic medicine, agriculture, biotechnology and have the changes in organisms generated by conventional potential to be used in pest control. breeding, gene editing, or natural mutation. What is a genome? The characteristics of all living organisms are determined BOX 1 by their genetic material and their interaction with the environment. An organism’s complete set of genetic HISTORIC SELECTION IN AGRICULTURAL CROPS material is called its genome which, in all plants, animals Some 6,000 – 10,000 years ago, Meso-American and microbes, is made of long molecules of DNA farmers began the drastic changes to a grass (deoxyribonucleic acid). The genome contains all the species called teosinte to become what is now genetic information needed to build that organism and known as maize. Through selecting and growing allow it to grow and develop.
    [Show full text]
  • Ethical Issues Regarding CRISPR-Mediated Genome Editing Zabta Khan Shinwari1,2*, Faouzia Tanveer1 and Ali Talha Khalil1
    Ethical Issues Regarding CRISPR-mediated Genome Editing Zabta Khan Shinwari1,2*, Faouzia Tanveer1 and Ali Talha Khalil1 1Department of Biotechnology, Quaid-i-Azam University, Islamabad. 2Pakistan Academy of Sciences, Islamabad, Pakistan. *Correspondence: [email protected] htps://doi.org/10.21775/cimb.026.103 Abstract Introduction CRISPR/Cas9 has emerged as a simple, precise Te quest for introducing the site-specifc changes and most rapid genome editing technology. With in the DNA sequence began when DNA was frst a number of promising applications ranging from discovered. Progress in genome engineering tech- agriculture and environment to clinical therapeu- nologies began in 1990s now reaching to a highly tics, it is greatly transforming the feld of molecular advanced, easy, economical and sophisticated biology. However, there are certain ethical, moral method for editing genomes called CRISPR/Cas9. and safety concerns related to the atractive applica- CRISPR (Clustered Regularly Interspaced Palin- tions of this technique. Te most contentious issues dromic Repeats) technology does not arrive as a concerning human germline modifcations are the breakthrough technology for editing the genomes challenges to human safety and morality such as risk but other genome editing platforms like TALENS of unforeseen, undesirable efects in clinical appli- (Transcription Activator-Like Efector Nucleases) cations particularly to correct or prevent genetic and ZFN (Zinc Finger Nucleases) were in use for diseases, mater of informed consent and the risk of some time but have lost their popularity because exploitation for eugenics. Stringent regulations and of their complexity, expensiveness and time con- guidelines as well as worldwide debate and aware- sumption (Carroll and Charo, 2015; Doudna and ness are required to ensure responsible and wise use Charpentier, 2014; Hsu et al., 2014; Jinek et al., of CRISPR mediated genome editing technology.
    [Show full text]
  • DP-Genome-Editing-EN-Web
    Discussion paper focusing on the scientific relevance of genome editing and on the ethical, legal and societal issues potentially involved ISSUED BY THE ETHICS COUNCIL OF THE MAX PLANCK SOCIETY 1. Introduction and motivation for the discussion paper Christiane Walch-Solimena As an organization dedicated to fundamental research, the intense controversies have emerged around some of its Max Planck Society (MPG) is committed to pursue issues at applications. Thus, first experiments in human embryonic cells the very frontiers of current knowledge and bears a special re- have been performed already since 2015 in China1 intended to sponsibility to critically evaluate novel scientific developments. correct certain disease-causing mutations. These publications Such assessment includes both the scientific potential as well have set off discussions throughout the scientific community as the risks that may be faced if the scientific findings may be and beyond about the ethical and safety implications of this put into practice one day in the future. To this end, the MPG research. An International Summit on Human Genome Editing Ethics Council has been asked to assemble a working group focused on the future of human genome editing and convened to outline and discuss questions arising from a revolutionary by the US National Academy of Medicine, the UK’s Royal Socie- technology that in recent years has opened up unforeseen ty and the Chinese Academy of Sciences in December 2015 opportunities in the manipulation of genes and genomes: the voiced the need for an ongoing global forum. Statements on CRISPR-Cas9 gene editing and genome engineering technolo- ethical and societal questions of genome editing, also cover- gy.
    [Show full text]
  • Potential Issues with CRISPR Rnai and CRISPR Screening Approach
    ` siPOOLs and CRISPR: Partnering RNAi with gene editing Catherine Goh, Michaela Beitzinger, Andrew Walsh, Michael Hannus siTOOLs Biotech GmbH, Lochhamerstrasse 29A, 82152 Martinsried/Planegg, Germany With the discovery of CRISPR, gene editing has gained tremendous traction in the scientific community due to its ease of application and low cost. The rise in popularity mirrors that of the use of RNA interference (RNAi) in the early 2000s. Although RNAi is still widely used today for functional genomics, screening efforts are hampered by the lack of specificity of individual siRNAs, giving rise to wide- ranging off-target effects and hence, unreliable results. Many are therefore now turning to CRISPR which promises higher specificity and clearer phenotypes due to complete depletion of gene activity. Here we compare key features of both technologies, bringing to attention that complete gene knock-out also comes with its own set of challenges. Of particular importance is the phenomenon of adaptation, which has now been shown to occur in several gene knock-out models. The complete knock-out of a gene can incur compensatory effects not seen when a gene is transiently knocked-down. With siPOOLs, transient knock-down by RNAi can be specific and potent. The dose-dependent nature of RNAi-mediated gene knock-down also mimics pharmacological inhibition. In view of their complementary strengths and challenges it seems highly commendable to use both RNAi and CRISPR for a thorough investigation and understanding of gene function. With siPOOLs, a new and extremely specific RNAi reagent has now become available that will allow RNAi screening with dramatically reduced off-target effects.
    [Show full text]
  • The Quiet Forest 203 References 207
    Fiona M. F. McQueen Our minds possess by nature an insatiable desire to know the truth. Cicero How can you reveal the truth if those to whom you are speaking do not want to hear? David Walsh, Author. Seven Deadly Sins. My pursuit of Lance Armstrong I This book is sold subject to the condition that it will not, by way of trade or otherwise, be re-sold, hired out or otherwise circulated without the publisher’s prior written consent, in any form of binding or cover other than that in which it is published and without a similar condition, including this condition, being imposed on the subsequent purchaser. No part of this book may be reproduced by any process, stored in a retrieval system, or transmitted in any form without the prior permission of the author. Copyright 2017. Fiona McQueen ISBN: 9781872970 55 9 Published by Tross Publishing, P.O. Box 22 143, Khandallah, Wellington 6441, New Zealand. Printed by Your Books, 18 Cashew Street, Grenada North, Wellington, New Zealand Cover Design, layout and production: Clark Design [email protected] II ABOUT THE AUTHOR Fiona McQueen (MBChB, MD, FRACP) was born in Dunedin in 1958 and graduated from the University of Otago in Medicine in 1980. She has worked as a consultant rheumatologist within the New Zealand public health system for the last 26 years, mostly in the Auckland region, but more recently in Southland. She completed an MD in Immunology (University of Auckland) in 1996 and was made Professor of Rheumatology in 2009. She has had a distinguished international academic career and has been active in research and teaching.
    [Show full text]
  • Winter – 2014 ISSN 1175-043X Protect
    Winter – 2014 ISSN 1175-043X Protect Our mission: Working together to ensure New Zealand is protected from the adverse impacts of invasive species WHAT IS WEED WEAPON™? ™ Weed Weapon is the culmination of years of research and XPI™ formula moves rapidly up and down the plant causing development in New Zealand and around the world. The cutting edge the membranes of cells in treated weeds to breakdown, formulation combines a new, world leading active ingredient and some allowing water to leak out of cells. The glyphosate can then enter the cells more easily, killing the entire weed faster. Kiwi magic to create the most effective weedkiller on the market. This new XPI™ Technology has a completely unique mode of action to other 1 XPI™ weedkillers (glyphosate). breaks down cells faster, ™ allowing Weed Weapon kills: glyphosate Grasses and broadleaf weeds like oxalis, dandelion, thistle, dock, to enter. clover, fennel, hemlock, onehunga, chickweed, speedwell, daisy, 2 Special geranium, mallow, ivy, ragwort, bindweed, nettle, plantain, formulation inkweed, willow herb, wild herbs, pennyroyal, henbit, hypericum, XPI™ rapidly spreads 3 vetch, lucerne, lupin, trefoil, milkweed, wild brassica, forget-me- through the Photosynthesis not, borage, groundsel, catsear, oxtongue, wild parsley, wild weed down to is inhibited, carrot, hemlock and more. the roots. weed dies. Visit our website for more gardening tips and information kiwicare.co.nz Replant in 5 days. Biodegradable in soil. Sprayed area safe for children and pets once dry. Kiwicare, PO Box 15050, Ph: +64 3 389 0778 Christchurch, NZ kiwicare.co.nz Protect Winter 2014 Magazine of the New Zealand Biosecurity Institute Contents Click on the item below to jump to the article.
    [Show full text]
  • CRISPR Gene Editing Amber Dance Doudna of the University of California, Science Writer Berkeley, One of the Developers of the Technology
    CORE CONCEPTS CORE CONCEPTS Core Concept: CRISPR gene editing Amber Dance Doudna of the University of California, Science Writer Berkeley, one of the developers of the technology. Some of the first researchers to investigate CRISPR were food scientists at Just a few years ago, molecular biologists an accompanying bit of RNA called a guide Danisco USA Inc.; they were worried about hoping to alter the genome of their favorite RNA. Then, the cell’s own DNA repair ma- viruses infecting the Streptococcus strains organisms faced an arduous task and likely chinery typically takes over in one of two used to make yogurt and cheese (1). These weeks of genetic tinkering. Today, those different modes. In the first mode, it simply researchers observed that bacterial chromo- scientists can quickly destroy or edit a glues the two pieces back together, but im- somes contain oddly repetitive sequences gene with a new technology called CRISPR perfectly, so the leftover scar interrupts and called “clustered regularly interspaced short (clustered regularly interspaced short palin- disables the targeted gene. Or, in a second palindromic repeats,” or CRISPR. Between dromic repeat)/Cas9. kind of repair, the cell can copy a nearby those repeats are sequences from viruses that “It really opens up the genome of virtu- piece of DNA to fill in the missing sequence. infect bacteria. The microbes use the viral ally every organism that’s been sequenced By providing their own DNA template, sci- sequences as a mnemonic to remember past to be edited and engineered,” says Jill entists can induce the cell to fill in any de- invaders.
    [Show full text]
  • Gene Editing Research Review an Overview of Recent Gene Editing Research Publications Featuring Illumina® Technology TABLE of CONTENTS
    Gene Editing Research Review An Overview of Recent Gene Editing Research Publications Featuring Illumina® Technology TABLE OF CONTENTS 4 Introduction The CRISPR Locus and the Mechanism for CRISPR-Cas Technology 7 Applications of CRISPR-Cas9 Technology Research Applications in the Medical Field Agriculture and Environmental Science 13 Workflow and Specificity Overview of the Procedure On-Target Effects Specificity: What It Means and Why It Matters Method Development Integration of CRISPR-Cas9 Technology in a Research Workflow 24 Bibliography This document highlights recent publications that demonstrate the use of Illumina technologies in single-cell research. To learn more about the platforms and assays cited, visit www.illumina.com. For Research Use Only. Not for use in diagnostic procedures. An overview of recent publications featuring Illumina technology 3 INTRODUCTION Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas technology 1. Jinek M, Chylinski K, Fonfara I, Hauer M, is a revolutionary gene editing method in which a programmable RNA guides a Doudna JA and Charpentier E. A program- nuclease to find a specific target location in the genome. This simple approach mable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. replaces the laborious and expensive protein-based DNA editing techniques, such 2012;337:816-821. as the use of zinc finger proteins (ZNFs) or transcription activator–like effector 2. Gasiunas G, Barrangou R, Horvath P and Siksnys V. Cas9-crRNA ribonucleoprotein nucleases (TALENs). From the time of the initial publications describing its application complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc Natl Acad 1, 2 3, 4 for genome editing in both prokaryotic and eukaryotic cells, the use of CRISPR- Sci U S A.
    [Show full text]
  • Advanced Gene Editing: CRISPR-Cas9
    Advanced Gene Editing: CRISPR-Cas9 Updated December 7, 2018 Congressional Research Service https://crsreports.congress.gov R44824 Advanced Gene Editing: CRISPR-Cas9 Summary Scientists have long sought the ability to control and modify DNA—the code of life. A gene editing technology known as CRISPR-Cas9 offers the potential for substantial improvement over other gene editing technologies in that it is simple to use and inexpensive and has a relatively high degree of precision and efficiency. These characteristics have led many in the scientific and business communities to assert that CRISPR-Cas9 will lead to groundbreaking advances in many fields, including agriculture, energy, ecosystem conservation, and the investigation, prevention, and treatment of diseases. Over the next 5 to 10 years, the National Academy of Sciences projects a rapid increase in the scale, scope, complexity, and development rate of biotechnology products, many enabled by CRISPR-Cas9. Concomitant with the promise of potential benefits, such advances may pose new risks and raise ethical concerns. For example, a Chinese researcher recently claimed that he had created the first genetically engineered human babies. According to the researcher, he used CRISPR-Cas9 to disable a gene that will make it harder for the twin girls, who were born in November 2018, to contract human immunodeficiency virus (HIV). The as yet unsubstantiated claim has sparked outrage and ethical debates by the international scientific community and others. Prior use of CRISPR-Cas9 gene editing in human embryos was generally limited to non- viable embryos, in part, to address ethical concerns such as the fact that the genetic change would affect not only the immediate patient, but also future generations who would inherit the change.
    [Show full text]