Welcome to The Conference
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The conference addressed applications of genome editing in a variety of biological systems, featuring:
- A high-impact, packed day of talks, discussions and several hours of networking
- Oral presentations on latest developments in the field of genome editing by an international faculty of leading researchers from academia and industry
- Update on CRISPR patent wars
- Ethical debate
- Posters
- Trade exhibition
- Excellent networking opportunities, and a relaxed and friendly environment
Distinguished Keynote Speakers
We welcome proposals for oral (now closed) and poster presentation on new applications and experimental design of genome editing and gene modulation technologies to large-scale screening, pathway elucidation, disease modelling, therapy and relevant emerging technologies. We particularly encourage final year doctoral students and early/mid career postdoctoral researchers/junior group leaders to submit oral abstracts, as we believe this event will provide their research an excellent exposure, critical analysis by attending senior researchers and new career opportunities. If you are interested in presenting your research at this meeting please contact us at CRISPR@LPMHealthcare.com.
The conference will be co-hosted with our 4th annual Aptamers 2017 and 3rd annual Oligo 2017 Oxford.
Speakers and Agenda
Morning Session: Chairs Drs Ben Davies & Mark Behlke
0920: Dr Ayal Hendel
Senior Lecturer, Bar-Ilan University, Israel
TITLE: Tuning the CRISPR Code: Chemically Modified RNAs Enhance CRISPR Genome Editing in Human Primary Cells
ABSTRACT: Targeted genome editing with engineered nucleases is increasingly allowing researchers to introduce precise sequence modifications at almost any site within the genome with nucleotide resolution. CRISPR/Cas-mediated genome editing relies on guide RNAs to direct site-specific DNA cleavage mediated by the Cas endonuclease. In this study, we chemically synthesized 100-nucleotide single guide RNAs (sgRNAs)…
0940: Dr Qingzhou Ji
Principal Scientist, MilliporeSigma/Merck, USA
TITLE: High Specificity and Enhanced HDR Efficiency CRISPR/Cas9 for Precision Genome Engineering
ABSTRACT: CRISPR (Clustered regularly interspaced short palindromic repeats) Cas9 nucleases have revolutionized the field of genome editing enabling unprecedented efficiency of gene targeting in a vast array of cell types and organisms. The potential therapeutic applications of this technology are restrained by low HDR (Homology directed repair) efficiency and off-target effects…
1000: Dr Mark Behlke (Keynote speaker)
Chief Scientific Officer, Integrated DNA Technologies, USA
TITLE: Genome editing using recombinant AsCpf1 and chemically modified crRNAs as a RNP complex
ABSTRACT: The Class 2 CRISPR/Cas system Cpf1 (Acidaminococcus sp.) is capable of mediating genome editing in mammalian cells. A single 41-44 base crRNA serves as the targeting guide for AsCpf1 comprising a 20 base constant region (loop domain) and a 21-24 base target-specific protospacer domain. In contrast to SpyCas9 sites which are identified by an ‘NGG’ protospacer adjacent motif (PAM) recognition sequence…
1030: Dr Houda Hallay
Coordinator of European Operations, Advanced Analytical, Germany
TITLE: Automating CRISPR mutation detection and zygosity determination
ABSTRACT: While CRISPR gene editing is rapidly advancing, becoming more economical and efficient, protocols to identify CRISPR mutations and determine the zygosity of these mutations are time consuming involving costly sequencing steps. To overcome this limitation, many researchers are turning to heteroduplexing and enzymatic mismatch cleavage assays to rapidly screen for mutated lines…
1050: Refreshments and networking – Exhibitors: Merck, IDT DNA, Advanced Analytical, Lonza, Dynamic Biosciences
1130: Dr Annaleen Vermeulen
Senior Scientist, Dharmacon/GE Healthcare, CO, USA
TITLE: Identification of genes involved in cell cycle regulation using a high-content, arrayed CRISPR RNA screening approach
ABSTRACT: Gene knockout using a CRISPR-Cas9 system has dramatically transformed biological research and enabled new types of genome-scale screening tools, including pooled lentiviral sgRNA libraries and arrayed synthetic crRNA libraries. Pooled sgRNA screening is limited by the requirements for assays that allow for enrichment or depletion in a cell population and deconvolution by next-generation sequencing…
1150: Dr Ben Davies
Transgenic Core Head and Group Leader, The Wellcome Trust Centre for Human Genetics, University of Oxford, UK
TITLE: Using CRISPR/Cas9 to probe the function of regulatory DNA
ABSTRACT: Site-specific nucleases, such as CRISPR/Cas9, have had an enormous impact on functional gene analysis, as we are now able to manipulate the genome of model systems with unprecedented speed and accuracy. The technology is frequently used to address the protein coding genome for the generation of knock-outs or models in which disease-associated variations in protein sequence have been knocked-in to the genome. It is becoming clear that the non-coding genome plays an important role in influencing disease susceptibility…
1210: Professor William James (Keynote speaker)
Professor of Virology and Pro-Vice-Chancellor, University of Oxford, UK
TITLE: Gene Editing in Human Pluripotent Stem Cells
ABSTRACT: The three techniques – induction of human pluripotent stem cells, CRISPR gene editing, and tissue differentiation in vitro – together provide a powerful new platform to investigate disease mechanisms and to evaluate potential therapies. Our lab investigates the role of tissue macrophages in immunodeficiency and neurodegenerative diseases, and have pioneered the development of these three techniques to answer previously intractable questions…
1240: Dr Philip Webber
UK and European Patent Attorney, Dehn Patent and Trade Mark Attorneys, Oxford, UK
TITLE: CRISPR: An update on the Patent Wars
ABSTRACT: A number of patents which cover the basic CRISPR technology have already been granted in the US and in Europe to Zhang’s group. Dounda’s group’s patent applications in the US and Europe are still pending, but they now have a granted UK patent. In the US Patent & Trademark Office, “interference” proceedings are underway which aim to resolve who owns the basic CRISPR technology in the US…
1300: Lunch and networking – Exhibitors: Merck, IDT DNA, Advanced Analytical, Lonza, Dynamic Biosensors
Afternoon Session: Chairs Drs Annaleen Vermeulen & Qingzhou Ji
1400: Dr Christopher Gyngell
Marie Sklodowska-Curie Fellow, The Uehiro Centre for Practical Ethics, University of Oxford, Oxford,UK
TITLE: The Ethics of Human Germline Gene Editing
ABSTRACT: Human Germline Gene Editing (GGE) has enormous potential both as a research tool and a therapeutic intervention. While other types of gene editing are relatively uncontroversial, GGE has been strongly resisted. In this talk, I analyze the ethical arguments for and against pursuing GGE. I argue there is a strong case for pursuing GGE for the prevention of disease…
1420: Dr Duncan Borthwick
Scientist, Dynamic Biosensors GmbH, Martinsried, Germany
TITLE: Modelling the Kinetics of Cas9 Interactions using switchSENSE Technology
ABSTRACT: A fuller understanding of the binding of Cas9 to target sequences will improve the design of assays and that of guide RNA sequences. switchSENSE® Technology utilises an electroswitchable biosurface and readily interchangeable nucleic acid target structures to generate real-time data on binding kinetics and activity. Assays can be performed without the need for heparin.
1440: Dr Lydia Teboul
Head of Molecular and Cellular Biology, MRC Harwell, Oxfordshire, UK
TITLE: Advances in the RGEN aided generation of complex alleles in mice
ABSTRACT: Mouse models are valuable tools to understand genes functions, genetic diseases and to develop and test new therapeutic treatments in vivo. The ability to introduce tailored modifications within the mouse genome is essential to generate them. The CRISPR/Cas system has brought new perspectives for the generation of mouse models in a more efficient and precise fashion, at reduced price, all within a shorter time scale…
1500: Dr Barry Rosen
Senior Principal Scientist, Discovery Sciences, AstraZeneca Cambridge, UK
TITLE: Precise Gene Editing to Empower Drug Discovery
ABSTRACT: Awaited
1520: Refreshments and networking – Exhibitors: Merck, IDT DNA, Advanced Analytical, Lonza, Dynamic Biosciences
1600: Dr Riley Doyle
CEO and Founder, Desktop Genetics Ltd, Hanbury Street, London, UK
TITLE: Accelerating clinical CRISPR through functional mapping of clinical genomic variant data to underlying disease biology
ABSTRACT: The wealth of data output from next-generation sequencing (NGS) technologies have led to tremendous progress in the study of the evolutionary origins of human genetic variation and its contribution to human disease. In particular, several consortia have been developed to map human genetic variation, providing genome-wide association studies and rare variant analyses in the context of common and rare diseases…
1620: Dr Xiaoping Li
Staff Scientist, The Wellcome Trust Sanger Institute, Cambridge, UK
TITLE: Genome Editing of the Pig Genome using TALENs and CRISPR
ABSTRACT: Genetically modified (GM) pigs hold great promise for agriculture, disease modeling, and xenotransplantation applications. However, porcine embryonic stem cells or induced pluripotent stem cells with germline chimeric ability have not been reported. GM- pig models are primarily generated by nuclear transfer following genome-editing in somatic cells. By taking the advantage of latest genome-editing tools such as TALENs and CRISPR…
1640: Miss Evgenia Sarrou
Graduate Research Student, Paul O’Gorman Leukaemia Research Centre, University of Glasgow, UK
TITLE: Altering the genome stability of haematopoietic cells using CRISPR/Cas9 technology
ABSTRACT: Haemopoiesis is a highly regulated process in which the most primitive cells have the ability to self-renew and give rise to the full repertoire of blood cells. Haemopoietic ageing has been associated with epigenetic alterations and genomic instability predisposing cells to the acquisition of mutations. In this study, we aimed to generate a haemopoietic model with altered genome stability using CRISPR/Cas9 technology…
1700: Dr Norah Fogarty
Postdoctoral Scientist, Niakan Laboratory, The Francis Crick Institute, London, UN
TITLE: CRISPR/Cas9 Targeting of Pou5f1/Oct4 in Mouse Zygotes
ABSTRACT: Much of our knowledge of mammalian preimplantation development comes from studies in the mouse as it is tractable model to perform genetic analyses. Indeed, while there are some gene expression patterns shared between the human and the mouse, we have reported that temporal expression dynamics of key developmental regulators and their co-expressed genes are largely distinct in human versus mouse…
1720: Dr Joey Riepsaame
Head of Genome Engineering Oxford, University of Oxford, Oxford, UK
TITLE: AAn introduction to the Genome Engineering Oxford (GEO) Facility – v2.0
ABSTRACT: AThe Genome Engineering Oxford (GEO) facility is a joint venture between the Dunn School of Pathology, DPAG, Biochemistry and Pharmacology. We offer advice on designing and implementing CRISPR-based gene editing experiments in a variety of eukaryotic systems, ranging from fly to human. We provide a CRISPR design and testing service to Oxford-based scientists for generating experimentally tested CRISPR/Cas9 plasmids and Cas9 ribonucleoproteins…
1740: Discussion and close
Posters
A CRISPR-scan assay for the functional characterisation of recurrent mutations in cancer
Erica Bello1, Qianxin Wu2, Andrea Pellagatti1, Jacqueline Boultwood1, Tudor A Fulga2
1Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, UK
2Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, UK
Recent advances in sequencing technologies have provided robust methods to gain insights into cancer genetics. However, the oncogenic role of many mutations identified by next generation sequencing and GWAS in cancer remains unknown. We developed a CRISPR-scan assay for the study of the oncogenic function of recurrent point mutations in cancer cells. We employed the CRISPR/Cas9 genome editing system to assess the effects of repairing mutations in cancer cells at the genome level. The chronic myeloid leukaemia…
Study of Type 2 Diabetes susceptibility loci with CRISPR/Cas9
Inês Cebola and Jorge Ferrer
Department of Medicine, Imperial College of London, Du Cane Road, London, W12 0NN, UK
Genome-wide association studies (GWAS) have identified over 100 loci carrying genetic variants associated with Type 2 Diabetes (T2D) susceptibility. However, the molecular mechanisms underlying most T2D genetic association signals remain elusive. It has been observed that transcriptional enhancers show enrichment for the presence of disease-associated SNPs, including T2D SNPs. In particular, T2D SNPs are enriched at enhancers that are active in pancreatic islets, a tissue that is central for diabetes aetiology. It is thus likely that…
Development of Genome Editing Surgery with Applicability to all Duchenne Muscular Dystrophy Patients
Helena Chaytow, Marc Moore, George Dickson, Linda Popplewell
Centre for Biomedical Sciences, Royal Holloway, University of London, UK
The mutations found in the dystrophin gene in Duchenne muscular dystrophy lead to a loss of the dystrophin protein. This protein is essential for normal muscle function, and so the loss in turn leads to muscle wasting. This project aims to introduce a smaller version of the dystrophin gene called microdystrophin into the mammalian genome, which produces a functional replacement for dystrophin. We targeted TALENs to the human AAVSI site, which is a conserved site in the mammalian genome known to allow efficient integration and stable expression of transgenes…
Tailored CRISPR/Cas9 Delivery System for Hard-to-Target Cell Lines
Solene Chartier, Leire Escudero-Ibarz, David Anderson, Thibault Laurent, Philippe Collin, Tom Henley
Horizon Discovery Group plc, Cambridge, UK
Modeling specific disease mutations in cell lines is essential to understand their biological consequence and the role they play in disease. Engineering cell lines with single nucleotide change, point mutation or reporter cassette insertion has been greatly facilitated with the adoption of CRISPR/Cas9 gene editing technology. However, one consistent element that has a significant impact on the success of that editing event when generating an isogenic cell line is the choice of parental cell line to be engineered…
An efficient plasmid-free and scalable CRISPR/Cas9 pipeline for epitope tagging in mammalian neural stem cells
Pooran Singh Dewari1, Ashley Tyrer1,2, Ashley Jacobi3, Mark Behlke3 and Steve Pollard1
1MRC Centre for Regenerative Medicine, SCRM Building, The University of Edinburgh, UK
2University of Bristol, UK
3Integrated DNA Technologies, USA
CRISPR-Cas9 technology has revolutionized precision genome manipulation across multiple organisms and cell types due to its adaptability, simplicity, and efficacy. Knock-in of small epitope tags into endogenous genes simplifies antibody-based assays, overcoming issues of specificity and sensitivity. Here, we demonstrate efficient and scalable CRISPR/Cas9-based epitope tag knock-in using mouse and human primary neural stem (NS) cell cultures. Three different methods of Cas9 delivery were compared: 1) transient expression through a plasmid, 2) recombinant Cas9 (rCas9) protein transfection, and 3) constitutive expression of Cas9 in cells…
Impact of genome variation on functional CRISPR screens and mapping of variant data to underlying disease biology
Ayokunmi Ajetunmobi, Leigh Brody, Victor Dillard, Neil Humphryes-Kirilov, Riley Doyle
Desktop Genetics, DESKGEN, London, United Kingdom
The wealth of data output from next-generation sequencing (NGS) technologies have led to tremendous progress in the study of human genetic variation and its contribution to human disease. Several consortia have been developed to map human genetic variation, producing genome-wide association studies and rare variant analyses in the context of common and rare diseases. However, many of these variant databases lack information on the function of these variants and their contribution to the pathophysiology of human diseases…
Footprint-free gene editing using Cas9/sgRNA RNP delivery and single-cell cloning of edited human iPS cells
Cornelia Hampe1, Montse Morell2, Tatiana Garachtchenko2, Hiro Matsumoto2, Michael Haugwitz2, and Andrew Farmer2
1Takara Bio Europe SAS, 78100 Saint-Germain-en-Laye, France
2Takara Bio USA, Inc., Mountain View, CA 94043, USA
While the introduction of CRISPR/Cas9 technology has made gene editing easier to achieve, obtaining single-cell clones of edited hiPS cells has been a major bottleneck. hiPS cells are traditionally grown and passaged as colonies. In order to obtain single cells for cloning purposes, the cells must first be dissociated into a single-cell suspension, which often results in cell death or premature differentiation. In contrast to traditional methods, the Cellartis® DEF-CS™ 500 Culture System allows culturing of hiPS cells in a monolayer and permits single-cell…
Comparative Analysis of CRISPR/Cas9 Delivery Methods
S Wakerell1, C Cieza-Borrella1, R Eeles1, 2, Z Kote-Jarai1
1The Institute of Cancer Research, London, UK
2The Royal Marsden Hospital NHS Trust, London, UK
CRISPR/Cas9 is rapidly gaining popularity as a genome editing tool, emerging as a potential alternative to TALEN and ZFN technology. Principally a quick, versatile and fairly simple system, however, various caveats can delay successful isolation of desired clones. We aimed to develop a robust method for gene KO in LNCaP and RWPE-1 cell lines by comparing different CRISPR/Cas9 delivery methods. RAD51B and RAD23B genes were knocked out using customized all-in-one vectors (SIGMA). Cells were transfected 24hrs post-seeding by electroporation…
Sponsors
Gold Sponsor, Exhibitor and Programme Partner
Merck is the preeminent life science company, supplying Sigma-Aldrich brand gene editing products and services. With over 300,000 products, including CRISPR reagents, whole genome screening libraries (including Sanger arrayed and GeCKO pooled libraries), validation services and technical expertise, Merck has committed to solving the toughest problems in life science.
Website: www.sigma-aldrich.com.
Gold Sponsor and Exhibitor
Advanced Analytical Technologies Inc. (AATI) develops, manufactures and markets high-throughput, fully-automated nucleic acid and genetic analysis systems. The company’s products have both commercial and research applications and are designed to improve processes within the molecular diagnostics, pharmaceutical, life science, agricultural and biofuels industries. The company’s product portfolio includes instruments for the parallel analysis of biomolecules, DNA, RNA, genomic DNA, double-stranded DNA, gene editing (CRISPR/Cas9), using capillary electrophoresis (CE) with fluorescence detection or UV absorbance. The company’s flagship product, the Fragment Analyzer, is recognized as the best-in-class, multi-channel, automated fluorescence-based CE detection system for the simultaneous analysis of the quantity and quality of nucleic acids, including: dsDNA fragments, gDNA, NGS fragments and RNA (total and messenger) and microsatellites (SSR). Advanced Analytical Technologies, Inc. (AATI) simplifies complex genomics workflows to accelerate research and discovery in pharmaceuticals, life science, biofuels, biotechnology and healthcare. The company has facilities in Ames, Iowa, USA and Heidelberg, Germany. We support customers through a global network of distributors and support centers.
Gold Sponsor and Exhibitor
Integrated DNA Technologies (IDT) is a leader in the manufacture and development of products for the research and diagnostic life science market. The largest supplier of custom nucleic acids in the world, IDT serves academic research, biotechnology, and pharmaceutical development communities.
IDT products support a wide variety of applications, including next generation sequencing (NGS), DNA amplification, SNP detection, microarray analysis, expression profiling, gene quantification, and synthetic biology. Platform-independent NGS products and services are available in addition to DNA and RNA oligonucleotides, qPCR assays, siRNA duplexes, and custom gene synthesis. Individually-synthesized xGen™ Lockdown™ Probes enable improved target capture. IDT also manufactures custom adaptors, fusion primers, Molecular Identifier tags (MIDs), and other workflow oligonucleotides for NGS. A TruGrade™ processing service is also available to reduce oligonucleotide crosstalk during multiplex NGS.
Serving over 80,000 life sciences researchers, IDT is widely recognized as the industry leader in custom oligonucleotide manufacture due to its unique capabilities. IDT pioneered the use of high throughput quality control (QC) methods and is the only oligonucleotide manufacturer to offer purity guarantees and 100% QC. Every oligonucleotide is analyzed by mass spectrometry and purified oligonucleotides receive further analysis by CE and HPLC. The company maintains an engineering division dedicated to advancing synthesis, processing technology, and automation. An in-house machine shop provides rapid prototyping and custom part design/control. Additionally, IDT offers unrivalled customer support, receiving approximately 100,000 calls annually with an average wait time of only 8 seconds.
A dedicated GMP manufacturing facility for molecular diagnostics provides oligonucleotides for In Vitro Diagnostic Devices (IVDs) or Analyte Specific Reagents (ASRs) for Laboratory-Developed Tests (LDTs). This manufacturing process is customer-defined and controlled, and facilitates progression from research to commercialization.
Bronze Sponsor and Exhibitor
Lonza provides the pharma market with the tools that life-science researchers use to develop and test therapeutics, beginning with basic research stages on to the final product release. Lonza’s bioscience products and services range from cell culture and discovery technologies for research to quality control tests and software that ensures product quality. Lonza Bioscience Solutions serves research customers worldwide in pharmaceutical, biopharmaceutical, biotechnology and personal care companies. The company delivers physiologically relevant cell biology solutions and complete solutions for rapid microbiology.
Lonza Cologne GmbH, Nattermannallee 1, Koeln, 50829, Germany
Phone: +49-221-99199-0, Fax: +49-221-99199-111
e-mail: scientific.support.eu@lonza.com
Exhibitor
Biophysical Analysis of Molecular Interactions with switchSENSE®
switchSENSE® is an automated biosensor chip technology that employs electrically actuated DNA nanolevers for the real-time measurement of binding kinetics (kON, kOFF) and affinities (KD). Interactions between proteins, DNA/RNA, and small molecules can be detected with femto-molar sensitivity. At the same time, protein diameters (DH) are analyzed with Angstrom accuracy and conformational changes as well as melting transitions (TM) can be measured using minimal amounts of sample.
The principles and applicability of three complementary measurement modalities provided by switchSENSE® will be introduced in this talk: Fluorescence Proximity Sensing, Molecular Ruler Measurements, and Switching Dynamics Measurements. In addition to standard workflows we’ll discuss unique possibilities for the functionalization of the sensor surface, i.e. the electrical adjustment of ligand densities and the precise assembly of different ligands on bifunctional nanolevers.
Application examples from drug development, quality control, and fundamental research will be discussed, including:
- – Analysis of complex binders: high-affinity antibodies and bispecific antibody formats
- – Small molecule induced conformational changes in proteins including dimers and aggregates
- – RNA/DNA binding proteins including riboswitches and aptamers
- – ‘large’ object binding including liposomes and virus-like particles
- – nucleic acid : nucleic acid methodologies for genotyping, antisense and SNP detection
- – Enzymatic activity of polymerases and CRISPR/Cas9
An introductory movie is presented here https://www.youtube.com/watch?v=amGw7txpGzM&autoplay=1
