Speakers and Agenda
DAY 1, 06th April 2016
13.00: Registration, welcome coffee, networking and exhibition
13.55: Welcome and housekeeping
Session 1: Chair Dr Gregory Davis
14.00: Opening KEYNOTE: Professor Emmanuelle Charpentier, Director, Max Planck Institute for Infection Biology, Berlin, Germany
14.40: Dr Mark Behlke, Chief Scientific Officer, Integrated DNA Technologies, USA
Improved CRISPR editing using chemically-modified crRNA:tracrRNA complexes
15.10: Dr Annaleen Vermeulen, Senior Scientist, Dharmacon/GE Healthcare, Lafayette, CO, USA
Utilizing synthetic RNA for CRISPR-Cas9 arrayed screening
15.40: Refreshment break, networking, exhibition and posters
16.10: Dr Eric Paul Bennett, Associate Professor, Copenhagen Center for Glycomics (CCG), University of Copenhagen, Denmark
“Hit and seek”: Improved strategies for identifying the breaks in a precisely broken genome
16.40: Dr Stephen Hague, European Droplet Digital PCR Specialist, Bio-Rad Laboratories, UK
Ultra-sensitive Quantification of Genome Editing Events by Droplet Digital PCR (ddPCR)
17.10: Dr Kyle Luttgeharm, Application Specialist, Advanced Analytical Technologies Inc. 2450 SE Oak Tree Ct, Ankeny, IA, USA
Development of a high throughput screening protocol for rapid identification of mutated alleles
17.30: Dr Cornelia Hampe, Scientific Support Specialist, Takara Bio Europe, France
Gene editing with high efficiency and no additional footprint using novel CRISPR/Cas9 Gesicle technology
17.50: Technology Workshop 1 by Merck KGaA
18.20: Close of Day 1
DAY 2, 07th April 2016
08.45: Welcome to Day 2
Session 2: Chair Dr Mark Behlke
08.50: Dr Ben Davies, Transgenic Core Head and Group Leader, The Wellcome Trust Centre for Human Genetics, University of Oxford, UK
Efficient CRISPR/Cas9 genome engineering using embryos derived from Cas9 overexpressing transgenic mice
9.20: KEYNOTE: Dr Gregory Davis, Senior R&D Manager, MilliporeSigma, USA
Genome and Epigenome Modification with ZFNs and CRISPR/Cas Systems
10.00: Dr Chady Jaber, Thermo Fisher Scientific, UK
How to boost your CRISPR genome editing efficiencies
10.30: Refreshment break, networking, exhibition and posters
11.00: Dr Philip Webber, Patent Attorney, Dehns Patent and Trademark Attorneys, Oxford, UK
CRISPR – the Patent Wars
11.20: Dr Barry Rosen, Senior Principal Scientist, Discovery Sciences, AstraZeneca, UK (TBA)
11.50: Technology Workshop 2 by Merck KGaA
12.20: Technology Workshop 2 by Dr Barry Rosen and Colleagues
13.00: Lunch, networking, exhibition and posters
Session 3: Chair Dr Barry Rosen
14.00: KEYNOTE: Dr William C Skarnes, Senior Group Leader, The Wellcome Trust Sanger Institute, UK
Biallelic genome editing of human iPSCs at scale
14.45: Professor Richard Wade-Martins, Professor of Molecular Neuroscience & Principal Investigator, The Oxford Parkinson’s Disease Centre, University of Oxford, UK
Deciphering the molecular basis of neurodegeneration using human iPSC neurons
15.15: Dr Robin Ketteler, Group Leader, MRC Laboratory for Molecular Biology and Manager, Translational Research Resource Center, University College London, UK
Applications of Genome Editing and RNAi to Dissect Autophagy Signaling Pathways
15.45: Dr Christine Seidl, Research Associate, The Kennedy Institute of Rheumatology, University of Oxford, UK
Investigating functional microRNA target sites by CRISPR/Cas9 genome editing
16.15: Refreshment break, networking, exhibition and posters
17.00: Dr Tony Nolan, Senior Research Fellow, Imperial College London, UK
A CRISPR-based gene drive system to suppress populations of malarial mosquitoes
17.30: Dr Michal Minczuk, Group Leader, Mitochondrial Genetics, MRC Mitochondrial Biology Unit, Cambridge, UK
Manipulating the human mitochondrial genome with designer nucleases
18.00: Dr Aleksandar Vojta, Associate Professor, University of Zagreb, Faculty of Science, Zagreb, Croatia
Targeted CpG methylation using the modified CRISPR-Cas9 system
18.30: Close of Day 2
19.15: Networking Dinner (by prior booking or invitation only)
DAY 3, 08th April 2016
08.35: Welcome to Day 3
Session 4: Chair Dr Eric Paul Bennett
08.40: Dr Victor Dillard, Chief Operating Officer, Desktop Genetics, London, UK
Designing for success: the right CRISPR design strategies for the right experiment
09.00: Dr Lydia Teboul, Head of Molecular and Cellular Biology, MRC Harwell, UK
CRISPR-aided mutagenesis of the mouse genome
09.30: Dr Emmanouil Metzakopian, Career Development Fellow, Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK
Human and Mouse Genome Wide CRISPR-guide RNA Arrayed Libraries
10.00: Dr Fanny Decarpentrie, Postdoctoral Research Fellow, James TURNER group, Sex chrom in development & disease, The Francis Crick Institute, London, UK
New tool for new discoveries: CRISPR/Cas9 genome editing in marsupials
10.30: Refreshment break, networking, exhibition and posters
11.00: Dr Cedric Ghevaert, Senior Lecturer, Transfusion Medicine & Consultant Haematologist, University of Cambridge/Cambridge Blood Centre, UK
Platelet production in vitro for transfusion from human pluripotent stem cells: the added power of genome editing
11.30: Dr Cristina Fimiani, Graduate Researcher, SISSA, Trieste, Italy
RNA-therapeutics of gene haploinsufficiencies
11.50: Dr Hamid Dolatshad, Research Associate, RDM-Clinical Laboratory Sciences, John Radcliffe Hospital, Oxford, UK
Use of CRISPR/CAS9 to investigate the function of gene mutations found in myeloid malignancies
12.20: Discussion and Close
Poster
Silencing Aspergillus terreus yap1 gene to clarify its role in the regulation of lovastatin biosynthetic genes
Ailed Pérez-Sánchez, Roxana Miranda and Javier Barrios-González
Departamento de Biotecnología, Universidad Autónoma Metropolitana Unidad Iztapalapa UAM-I, C.P. 09340, México, D.F, México
ABSTRACT: Lovastatin (LOV) is a secondary metabolite, produced by Aspergillus terreus that lowers cholesterol levels in blood. Earlier studies of our group showed a link between reactive oxygen species (ROS) and LOV biosynthesis, since a ROS build up during production phase (idiophase) was detected. In a subsequent work we showed that ROS regulate LOV biosynthesis at a transcriptional level, although the mechanism is presently unknown. It is considered that stress response transcription factor(s) could be this link between ROS and the biosynthetic genes. A search for transcription factors binding sites in the promoter regions of the LOV genes: lovE and lovF, revealed putative sites for several oxidative stress response TFs including Yap1….
Development of tools for modulation of genes involved in protein glycosylation by genome editing
Luka Bočkor, Vanja Tadić, Paula Dobrinić, Dora Markulin, Aleksandar Vojta, Vlatka Zoldoš
Department of Molecular Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, 10000 Zagreb, Croatia
ABSTRACT: Glycosylation is one of the most important post-translational modifications of proteins contributing to a number of functions in protein biology, from protein folding and quality control to recognition events such as cell signalling and immune surveillance. Glycans have come into focus recently as contributors or cause of different human complex diseases. A recent GWAS study of the IgG glycosylation identified several genes with the IL6ST locus being among the top hits. IgG represents an excellent model glycoprotein because it has one glycosylation site at each heavy chain. Many of the structures present on IgG have been well defined with many important functional effects of alternative IgG glycosylation described…
Antisense Oligonucleotide Mediated Rescue of a Deep Intronic Point Mutation in OPA1
Tobias Bonifert1, Irene Gonzalez Menendez1, Matthis Synofzik2,3, Ludger Schöls2,3, Bernd Wissinger1
1 Molecular Genetics Laboratory, Institute for Ophthalmic Research, Centre for Ophthalmology, University of Tübingen, Germany
2 Department of Neurodegenerative Diseases, Hertie-Institute for Clinical Brain Research, University of Tübingen, Germany
3 Center for Neurodegenerative Diseases (DZNE), Helmholtz Association of German Research Centers, Tübingen, Germany
ABSTRACT: Inherited Optic Neuropathies (ION) represent an important cause of blindness in the European working-age population. Despite progress in mutation screenings and disease gene identification, a large number of all cases remain unsolved – partly due to exon-centered screening approaches. Recently we reported the discovery of four independent families with deep intronic point mutations (DIM) in OPA1 that cause mis-splicing of the pre-mRNA transcript by creating a cryptic 3’ splice site (ss). In accordance with the haploinsufficiency model, heterozygosity for a DIM leads to reduced levels of OPA1 protein which in turn drives disorganization of the patients’ mitochondria and provokes ION. As a rescue strategy we thought to prevent missplicing of the mutant pre-mRNA by applying 2’O-Methyl-Antisense Oligonucleotides (AONs)…
The utility of SmartFlare probes to detect heme oxygenase-1 transcript level during selection of CRISPR/Cas-generated HO-1 knockout cells
Maria Czarnek and Joanna Bereta
Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland
ABSTRACT: Knockout cells are invaluable research model in modern biological sciences. The ability to modify genomes in a precise and efficient fashion is needed to understand how genotype affects different physiological and pathological processes. Genome editing tools, including CRISPR/Cas9, enable researchers to modify virtually any locus across mammalian genome. However, modification efficiency may be the primary limitation. Cas9 can induce the formation of targeted double strand breaks in mammalian chromosomes. Subsequent indel formation via the nonhomologous end-joining (NHEJ) repair mechanism often introduces nonsense or frameshift mutations, which leads to nonsense-mediated degradation of mRNA. The aim of our project was to test whether SmartFlare probes…
Evaluation of CRISPR/Cas9 efficiency for muscle-specific Mir-378a targeting in vivo
Szymon Czauderna1, Maria Czarnek2, Mateusz Tomczyk1, Alicja Jozkowicz1 and Jozef Dulak1
1 Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow, Poland
2 Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow, Poland
ABSTRACT: CRISPR/Cas system was adopted to target eukaryotic DNA making it the most widely used method for genome editing due to its efficiency and simplicity. Our work aims to evaluate CRISPR/Cas efficiency of Mir-378a locus targeting in vivo using adeno-associated viruses as vectors for CRISPR/Cas delivery. sgRNA flanking Mir-378a were manually designed and cloned into pX601 plasmid (Addgene). sgRNA efficiency and plasmid performance were evaluated in Neuro-2a or C2C12 by Cel-I digestion, diagnostic PCR or western blot. Off-target prediction was performed using rgenome software. For dual sgRNA expression custom synthesized cassette was cloned into pX600 vector by Gibson assembly. For muscle-specific Cas9 expression…
Development of epigenetic CRISPR-Cas9 system for targeted methylation at specific CpG sites
Paula Dobrinić, Aleksandar Vojta, Vanja Tadić, Luka Bočkor, Marija Klasić, Petra Korać, Vlatka Zoldoš
Division of Molecular Biology, Department of Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, 10000 Zagreb, Croatia
ABSTRACT: DNA methylation is an important epigenetic mechanism involved in gene regulation. In mammals, DNA methylation mostly occurs at symmetrical CpG dinucleotides. Cytosine methylation of gene regulatory regions has usually been associated with gene silencing, but functional studies have been limited due to the lack of methods for targeted manipulation of methylation marks. To that end, we developed a flexible, easily programmable tool for targeted methylation at specific CpG sites in mammalian cells, based on the CRISPR-Cas9 system. We completely abolished the nuclease activity of Cas9 protein and added the DNA methyltransferase domain of human DNMT3A, using a short peptide linker…
Direct delivery of Cas9-sgRNA protein complex via cell-derived nanovesicles
Montse Morell, Tatiana Garachtchenko, Lily Lee, Thomas P. Quinn, Michael Haugwitz, and Andrew Farmer (to be Presented by Cornelia Hampe)
ABSTRACT: While CRISPR/Cas9 is a powerful technique for genome manipulation, two significant challenges remain: obtaining efficient delivery of Cas9 to all cell types and achieving fewer off-target effects. Recently, it has been demonstrated that genome editing via delivery of Cas9 protein is as effective as plasmid-based delivery, but with the added benefit of less off-target effects due to the short lifespan of the Cas9 protein in the cell (1). Here we report delivery of Cas9 protein using cell-derived nanovesicles called gesicles. Gesicles are produced by a mammalian packaging cell via co-overexpression of three components: a nanovesicle-inducing glycoprotein, Cas9…
Identifying somatosensory subtypes using CRISPR/Cas9-directed transgene integration
Donald P Julien, Alex W Chan and Alvaro Sagasti
Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles USA
ABSTRACT: The skin is a multimodal sensory organ, capable of detecting multiple types of touch stimuli (e.g. mechanical, thermal, and chemical). Underlying this sensory diversity are multiple populations of somatosensory axons that terminate as specialized sensory endings in the skin. The diversity of sensory termini in the skin and the how their organization is established during development, however, remains poorly understood. The embryonic zebrafish is an ideal model for investigating the development of somatosensory neurons because they can be imaged in live animals. However, differentiating individual sensory subtypes remains a challenge due to a paucity of transgenic reporters…
Engineering the mouse genome using CRISPR/Cas9 technology
Ruairidh King, Joffrey Mianné, Rachel Fell, Adam Caulder, Gemma Codner, Marina Maritati, Martin Fray, Wendy Gardiner, MLC Microinjection team, Sara Wells, and Lydia Teboul
The Mary Lyon Centre, MRC Harwell, HSIC, Oxon, OX11 0RD, UK
ABSTRACT: Mouse models are valuable tools to understand gene 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 these models of human diseases. The recently developed CRISPR/Cas9 system as genome engineering tool has brought new perspectives for the generation of mouse models in a precise fashion, at reduced price, and within a shorter time scale. Here we will report the use of the CRISPR/Cas9 technology at the Mary Lyon Centre, MRC Harwell, to engineer the mouse genome through different methods…
Candidate testing of axonal sprouting genes in a cortical circuit induced by limb overuse after stroke
Esther H. Nie1, Giovanni Coppola2, Riki Kawaguchi2, S. Thomas Carmichael1
1 Dept. Neurology, UCLA David Geffen School of Medicine, Los Angeles, CA
2 Depts of Psychiatry and Biobehavioral Sciences, UCLA David Geffen School of Medicine, Los Angeles, CA
ABSTRACT: Stroke survivors worldwide are left with permanent sensorimotor and cognitive disabilities for which there are no medical treatments. However, a behavioral paradigm of limb overuse after stroke, known as constraint-induced movement therapy (CIMT), has been shown in clinical trials to result in significant motor improvements. The goal of the current project is to understand how specific gene systems drive this important neural repair process. We have previously mapped a unique connection to premotor cortex upon post-stroke limb overuse. We find that CIMT-like limb overuse drives the activity-dependent formation of connections between premotor cortex (PMC) and retrosplenial cortex (RSC), a brain area involved in spatial learning. The finding was significant and replicated across two independent cohorts (p<0.05, Hotelling’s T test)…
Functional multiparameter analysis of double-strand breaks and associated biomarkers during genome editing
Lysann Sauer1, Stefan Rödiger1, Jens Schneider1, Peter Schierack1, Dirk Roggenbuck1,2 and Christian Schröder1
1 Faculty of Natural Sciences, Brandenburg University of Technology Cottbus – Senftenberg, Senftenberg, Germany
2 Medipan GmbH, Dahlewitz/Berlin, Germany
ABSTRACT: A powerful and highly precise technology, adapted from the microbial adaptive immune system, is enabling the systemic analysis of the mammalian genome. The CRISPR-associated RNA-guided endonuclease Cas9 induces target-specific double-strand break (DSB) thereby activating DNA repair pathways such as nonhomologous end joining (NHEJ) or homology-directed repair (HDR). The delivery of the CRISPR/Cas9 components and potential off-target mutagenesis are of concern and require further elucidation. This is of particular importance for clinical applications and personalized ; therefore, we used our automatized imaging platform…
Involvement of lncRNAs in muscle differentiation
Rossella Tita1, Monica Ballarino1, Andrea Cipriano1, Fabio Desideri1 and Irene Bozzoni1,2,3
1 Dept of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, Rome, Italy
2 Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Rome, Italy
3 Institute Pasteur Fondazione Cenci-Bolognetti and IBPM, Sapienza University of Rome, Rome, Italy
ABSTRACT: In recent years, it has been discovered that genomes of multicellular organism are characterized by the pervasive expression of different types of non-coding RNAs (ncRNA) and among these, long non-coding RNAs (IncRNAs) are included. A transcriptome analysis performed during in vitro murine muscle differentiation allowed us to identify new lncRNAs differentially expressed during myogenesis. These transcripts were classified on the basis of their expression in proliferating versus differentiated conditions, muscle-restricted activation and subcellular localization. We are now focusing on the characterization of a nuclear lncRNA, lnc-405, which is up-regulated in myotubes and conserved in human. To dissect its role in vivo we are using the CRISPR Cas9 strategy…
GEGM 2016 Sponsors
Gold Sponsor, Exhibitor and Programme Partner
Sigma-Aldrich, now a part of Merck, is a global supplier of leading RNA interference (RNAi) tools including siRNA and the comprehensive MISSION® shRNA library through participation in The RNAi Consortium (TRC). As a key licensed supplier of siRNA and expert in high quality custom DNA and RNA oligonucleotide synthesis, our goal is to provide innovative products to meet your functional genomics and gene silencing research needs. The MISSION shRNA library of The RNAi Consortium, developed by the Broad Institute of MIT and Harvard, is a pre-cloned collection targeted against annotated human and mouse genes enabling scientists to elucidate gene function. For further details of the above products please visit www.sigma-aldrich.com.
Sigma-Aldrich, a leading Life Science and High Technology company focused on enhancing human health and safety, manufactures and distributes more than 200,000 chemicals, biochemicals and other essential products to more than 1.4 million customers globally in research and applied labs as well as in industrial and commercial markets. With three distinct business units – Research, Applied and SAFC Commercial – Sigma-Aldrich is committed to enabling science to improve the quality of life. The Company operates in 37 countries, has more than 9,000 employees worldwide and had sales of $2.6 billion in 2012. For more information about Sigma-Aldrich, please visit its website at www.sigma-aldrich.com.
Gold Sponsor and Exhibitor
Bio-Rad Laboratories has played a leading role in the advancement of scientific discovery for 60 years by providing a broad range of innovative tools and services to the life science research and clinical diagnostic markets. Bio-Rad’s Life Science Group develops, manufactures, markets and supports a wide range of laboratory instruments, apparatus and consumables used for research in genomics, proteomics, cell biology and food safety.
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Meanwhile, researchers studying disease are now using ddPCR for more powerful screening of genome editing and efficient CRISPR validation. Droplet Digital PCR is also showing great promise or diagnostics, making liquid biopsy a reality through reliable detection of circulating tumour DNA and cell-free microRNA.
Gold Sponsor and Exhibitor
Precise, efficient gene sequence targeting and modification technologies from ThermoFisher Scientific.
From precision genome editing and gene modification technologies to high-efficiency delivery systems, we have developed a broad range of solutions to help you create the modified genes, expression systems, and stable cell lines you need for your research—from culturing cells to modification, detection, and analysis. Our technologies—including GeneArt® TALs andCRISPRs—enable you to reliably modify genomic sequences and analyze the phenotypic outcomes. In addition, we can deploy a combination of advanced technologies to produce a custom construct or cell line that meets your specifications. Balancing performance and cost, our cell engineering portfolio is built on 20 years of industry-leading innovation and can grow with your research needs.
Silver 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.
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Bronze Sponsor and Exhibitor
Takara Bio Europe and Clontech Laboratories are members of the Takara Bio Group, a leading supplier of tools for life scientists worldwide. Through our brand names TAKARA and CLONTECH we develop innovative technologies in the fields of Stem Cells and Epigenetics, Molecular and Cell Biology, and Gene/Protein Function. Key products include SMARTer™ cDNA synthesis kits for next generation sequencing, the innovative In-Fusion® HD Cloning Plus System, high performance PCR/qPCR reagents, Tet-regulated gene expression systems, Living Colors® Fluorescent Proteins, as well as a broad choice of viral vectors/particles and transduction tools.
Our recently launched Guide-it™ kits are excellent tools for genome engineering using CRISPR/Cas9 technology. The Guide-it™ portfolio includes kits for efficient in vitro production and screening of single guide RNAs (sgRNAs) as well as Mutation Detection and Indel Identification kits for monitoring the efficiency of genome editing.
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Bronze 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.
Bronze Sponsor and Exhibitor
Eupheria Biotech GmbH is a young, dynamic and fast growing company, which was founded in 2010 as a spin-off of the Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG) in Dresden, introducing highly specific and efficient silencers: esiRNAs!
In the meantime, RNA interference screens with esiRNAs have proven to be a very powerful technology to unravel gene function in basic and applied research. The specificity and cost-effectiveness of esiRNAs have revolutionized drug discovery.
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