FRONTIERS IN PLANT SCIENCE 08/11/16 Engineering Plant Immunity: Using CRISPR/Cas9 to Generate Virus Resistance. Introduction In the context of the rapidly growing global population, food security has emerged as one of the major challenges facing our generation (Cheeseman, 2016).
The global population has increased by 60%, but per capita production of grains has fallen worldwide in the last 20 years (Suweis et al., 2015). If the population growth rate, which is 1.13 percent per year for 2016 persists, the world population will double again within a mere 50 years, and it is estimated that food production will need to at least double till 2050 to meet demand (Suweis et al., 2015). Increases in food production per unit of land have not kept pace with increases in population and cropland area per capita has fallen by more than half since 1960 (Cheeseman, 2016). Plant Viruses Agriculture worldwide is threatened by abiotic (heat, drought, frost, salinity, etc.) and biotic stresses (insect pests, fungi, bacteria, viruses, etc.). Geminiviruses FIGURE 1. Virus Control Strategies. PEERJ 09/05/16 Genome editing weds CRISPR: what is in it for phytoremediation?
Journal of Insect Physiology Volume 98, April 2017, CRISPR/Cas9 in insects: Applications, best practices and biosafety concerns. HORIZON MAGAZINE (EU) 18/05/17 Can CRISPR feed the world? By 2040, there will be 9 billion people in the world.
‘That’s like adding another China onto today’s global population,’ said Professor Sophien Kamoun of the Sainsbury Laboratory in Norwich, UK. Prof. Kamoun is one of a growing number of food scientists trying to figure out how to feed the world. NATURE 16/05/17 Geneticists enlist engineered virus and CRISPR to battle citrus disease. Joe Raedle/Getty Tangerine groves in the southern United States are vulnerable to a disease known as citrus greening.
Fruit farmers in the United States have long feared the arrival of harmful citrus tristeza virus to their fields. But now, this devastating pathogen could be their best hope as they battle a much worse disease that is laying waste to citrus crops across the south of the country. The agricultural company Southern Gardens Citrus in Clewiston, Florida, applied to the US Department of Agriculture (USDA) in February for permission to use an engineered version of the citrus tristeza virus (CTV) to attack the bacterium behind citrus greening.
This disease has slashed US orange production in half over the past decade, and threatens to destroy the US$3.3-billion industry entirely. The required public comment period on the application ended last week, and the USDA will now assess the possible environmental effects of the engineered virus. NATURE SCIENTIFIC REPORTS 03/05/17 Construction of a highly efficient CRISPR/Cas9-mediated duck enteritis virus-based vaccine against H5N1 avian influenza virus and duck Tembusu virus infection. Rapid generation of recombinant virus C-KCE-HA/PrM-E encoding HA and PrM-E based on CRISPR/Cas9 mediated gene editing To exclude the adverse effects of viral gene modification on viral replication, the two gene junctions UL27/UL26 and US7/US8 of C-KCE, which our recent studies have proven to be suitable for foreign gene insertion13, 28, were selected as the target regions for recombinant C-KCE generation (Fig. 1A and B).
Schematic illustration of the novel candidate trivalent vaccine C-KCE-HA/PrM-E development. (A) Full-length of the attenuated commercial DEV vaccine strain (C-KCE). (B) Two portions of the genome C-KCE expanded to show the UL27, UL26, US7, and US8, and the gene junction regions are depicted. ECDC 03/05/17 ECDC assesses risk of ‘do-it-yourself’ CRISPR gene engineering kit contaminated with pathogenic bacteria. On 24 March 2017, German authorities reported the contamination of a ‘do-it-yourself’ (DIY) Bacterial Gene Engineering CRISPR (Clustered regularly interspaced short palindromic repeats) kit with pathogenic bacteria (risk group 2), including some that are multidrug-resistant with production of Extended Spectrum Beta-Lactamase (ESBL).
The kits are produced in the United States and sold over the internet, targeting non-professional users who want to study biology and life science using similar biotechnology engineering tools found in laboratory settings. In its risk assessment published today, ECDC identifies the risk of infection for users of the kits unaware of the contamination with pathogenic agent as low, as the manipulation of the kit does not involve percutaneous injury-prone manipulations. However, infection resulting from the contamination of broken skin or mucous membranes may occur, even though the kit recommends and provides disposable gloves. ARS USDA - 2016 - Projet de recherche : CONTROL OF WHITE SPOT SYNDROME VIRUS (WSSV) OF SHRIMP USING GENOME EDITING.
Performing Department Research & Development Non Technical Summary No drugs or treatmentsare available to combat viral diseases in shrimp such as the white spot syndrome disease (WSD) caused by white spot syndrome virus (WSSV).
Viral diseases can rapidly spread in shrimp stocks and threaten the viability of the US and global shrimp aquaculture industry. While management and good biosecurity are part of the solution, having a direct method to respond to an outbreak or to assure incoming shrimp are clear of virus would be a strong additional tool for the industry.The use of genome editing technology (such as CRISPR/Cas)to attack the invading organism rather than to modify the genome of the host contrasts with most current applications of this new technology (i.e., targeting the genome of the host organism). Animal Health Component Research Effort Categories Basic. Theriogenology. 2016 Jul 1;86(1):160-9. New insights and current tools for genetically engineered (GE) sheep and goats.
Nucleic Acids Res. 2013 Nov;41(20):e188. Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice. Mol Plant Pathol. 2016 Jan 25. Development of broad virus resistance in non-transgenic cucumber using CRISPR/Cas9 technology. GEN 03/01/17 Genetic Engineering and Crops: The CRISPR Conundrum. Public Policy Issues In both the U.S. and the EU, transgenic genetically modified (GM) crops must be approved by regulatory agencies before they can be marketed.
Both the U.S. and EU have mandatory GM labeling requirements, though the structures of the U.S. law, which was passed in July, are far less imposing and rigorous than that of the EU. Regulation of transgenic GM crops by the USDA is rooted in the Plant Protection Act (PPA), which broadly gives the USDA authority to regulate plant pests or noxious weeds, including GM organisms that are, or have the potential to be, plant pests.
However, the USDA has signaled that genetically engineered crops that do not contain “foreign” DNA are not considered to be GM crops, and thus do not require USDA regulation. EMBO - NOV 2015 - No time to waste — the ethical challenges created by CRISPR. Nucleic Acids Res. 2013 Nov;41(20):e188. Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice. GENOME BIOLOGY 01/02/17 Single Cas9 nickase induced generation of NRAMP1 knockin cattle with reduced off-target effects. Prediction and selection of the targeting locus and sgRNAs Considering the potential synergistic effects of neighboring genes, we used an intergenic region between the fascin actin-bundling protein 1 gene (FSCN1) and the actin beta gene (ACTB) on chromosome 25 (positions 40,631,870–40,632,430; 561 bp long) as the potential gene target region (Additional file 1: Supplemental dataset S1).
Housekeeping genes in the FSCN1-ACTB (F-A) locus (Fig. 1a) have relatively steady expression levels across various tissues and thus exogenous gene silencing resulting from chromatin inactivation might be avoided. We used the open-source website ZiFiT [21, 22, 23] ( and identified a total of 80 target sites in this locus. These sites either ended with NGG or started with CCN on the reverse strand (i.e. the protospacer adjacent motif (PAM))  (Additional file 2: Supplemental dataset S2). PHYS_ORG 31/01/17 Tuberculosis-resistant cows developed for the first time using CRISPR technology. CRISPR/Cas9 gene-editing technology has been used for the first time to successfully produce live cows with increased resistance to bovine tuberculosis, reports new research published in the open access journal Genome Biology.
The researchers, from the College of Veterinary Medicine, Northwest A&F University in Shaanxi, China, used a modified version of the CRISPR gene-editing technology to insert a new gene into the cow genome with no detected off target effects on the animals genetics (a common problem when creating transgenic animals using CRISPR). Dr Yong Zhang, lead author of the research, said: "We used a novel version of the CRISPR system called CRISPR/Cas9n to successfully insert a tuberculosis resistance gene, called NRAMP1, into the cow genome. We were then able to successfully develop live cows carrying increased resistance to tuberculosis. MOTHERBOARD 21/11/16 Obama’s Science Advisors Are Worried About Future CRISPR Terrorism. Last week the President’s Council of Advisors on Science and Technology (PCAST), which consists of 18 scientists and policy experts in various disciplines, issued a letter to President Obama on the potential emergence of new forms of bioterrorism.
“While the ongoing growth of biotechnology is a great boon for society, it also holds serious potential for destructive use by both states and technically-competent individuals with access to modern laboratory facilities,” the PCAST members wrote. “Molecular biologists, microbiologists, and virologists can look ahead and anticipate that the nature of biological threats will change substantially over the coming years. The U.S. Government’s past ways of thinking and organizing to meet biological threats need to change to reflect and address this rapidly-developing landscape.” As detailed in the letter, the US approach to biological threats for the last two decades has focused on a “dangerous subset of known human and agricultural pathogens.”
BERKELEY_EDU 24/01/17 CRISPR research institute expands into agriculture, microbiology. An initiative launched two years ago by UC Berkeley and UC San Francisco to use CRISPR-Cas9 gene editing to develop new disease therapies is expanding into research on the planet’s major crops and poorly understood microbiomes, with plans to invest $125 million in these areas over the next five years. The funds will not only boost support for biomedical research, but also allow the renamed Innovative Genomics Institute to explore the potential of gene editing in the globally important areas of agriculture and microbiology, and fund projects focused on the social and ethical implications of editing human, animal and plant genomes.
“The CRISPR-Cas9 technology, which is only four years old, is improving by leaps and bounds and has already altered the way doctors approach disease and scientists do research,” said IGI executive director Jennifer Doudna, a professor of molecular and cell biology and Howard Hughes Medical Institute investigator at UC Berkeley. GEN 03/01/17 Genetic Engineering and Crops: The CRISPR Conundrum. Future Microbiology December 2016 CRISPR–cas loci profiling of Cronobacter sakazakii pathovars. SCIENTIFIC REPORTS 17/08/16 Efficient Genome Editing in Apple Using a CRISPR/Cas9 system. Cloning of the apple PDS gene In order to test whether the CRISPR/Cas9 system can induce mutation effectively in the apple genome, we selected the apple phytoene desaturase (PDS) gene, which is required for chlorophyll biosynthesis5 as a target gene.
In other plant species, pds mutant plants show an albino phenotype5, and have served as a model for CRISPR/Cas9-mediated targeted gene editing4,6,7,8. A 4801-bp fragment corresponding to the partial apple PDS genome region was cloned (accession no. LC101839) from rootstock cultivar ‘JM2’. Comparison of reported apple PDS ESTs and PDS ESTs from other plant species showed that the cloned region spanned from the middle of the third exon to the middle of the ninth exon of the apple PDS gene (Fig. 1a). PHYSORG 29/11/16 Enhanced CRISPR lets scientists explore all steps of health and disease in every cell type. Wellcome Trust Sanger Institute and University of Cambridge researchers have created sOPTiKO, a more efficient and controllable CRISPR genome editing platform. Today, in the journal Development, they describe how the freely available single-step system works in every cell in the body and at every stage of development. This new approach will aid researchers in developmental biology, tissue regeneration and cancer.
Two complementary methods were developed. sOPiTKO is a knock-out system that turns off genes by disrupting the DNA. sOPTiKD is a knock-down system that silences the action of genes by disrupting the RNA. Using these two methods, scientists can inducibly turn off or silence genes, in any cell type, at any stage of a cell's development from stem cell to fully differentiated adult cell. The body contains approximately 37 trillion cells, yet the human genome only contains roughly 20,000 genes. Share Video undefined. FRONTIERS IN PLANT SCIENCE 08/11/16 Engineering Plant Immunity: Using CRISPR/Cas9 to Generate Virus Resistance.
BIOTECHNIQUES - NOV 2016 - CRISPR: MODIFYING THE LIFE SCIENCE LANDSCAPE. BioTechniques, Vol. 61, No. 5, November 2016, pp. 225–231 Abstract From lampreys to human stem cells, the CRISPR/Cas9 system is challenging our notions of what is possible with genome editing. Nathan Blow talks to researchers pushing the boundaries of CRISPR/Cas9 technology to expand our understanding of biology. Three years ago, University of Colorado Boulder researcher Daniel Medeiros was at a crossroads in his study of vertebrate evolution. He was interested in a critical period in the history of vertebrate physiology: the transition from jawless vertebrates to jawed vertebrates. “I was actually thinking about winding down the lamprey work,” says Medeiros. Previously, Medeiros’ lab had made some headway using pharmacological inhibitors and morpholinos to study very-early gene function in lamprey, but there are a limited number of chemicals available, and he was really interested in looking at later developmental processes, in particular the formation of the head skeleton.
AGRICULTURE ENVIRONNEMENT 07/11/16 CRISPR/Cas9 au service des concombres. EFSA 12/04/16 Literature review of baseline information on non-coding RNA (ncRNA) that could support the food/feed risk assessment of ncRNA-based GM plants. Molecular Biotechnology September 2013, Volume 55, Issue 1, pp 87-100 Non-coding RNAs in Crop Genetic Modification: Considerations and Predictable Environmental Risk Assessments (ERA) EUREKALERT 11/09/14 Crop improvement and resistance to pathogens benefits from non-coding RNA studies. With the rise of emerging economies around the world and a concomitant upgrade of health care systems, the global population has been rapidly expanding.
As a consequence, worldwide demand for agricultural products is also growing. Crops now provide food and the other important resources for seven billion humans. Food supplies are primarily based on such crops as wheat, maize, rice and vegetables. But as the area of arable land and of cultivated land continues to decline, the future ability to meet the world's food security needs has come under a cloud of uncertainty. Meanwhile, the use of pesticides and fertilizers has triggered long-term adverse effects on the environment, and has presented a serious threat to human health.
Proponents of new technologies including DNA recombination have promised a new green revolution, with genetically modified crops featuring transgenes achieving targeted traits including improved quality, increased production, and resistance to pests and stress. BUNDESMINISTERIUM FUR GESUNDHEIT (AT) - DEC 2013 - New plant breeding techniques RNA-dependent DNA methylation, Reverse breeding, Grafting. ISIS 16/06/14 Non-Coding RNA and Evolution of Complexity. ENSIA_COM 28/01/16 CRISPR is coming to agriculture — with big implications for food, farmers, consumers and nature.
NPR 15/04/16 Will Genetically 'Edited' Food Be Regulated? The Case Of The Mushroom. Biotechnology Advances Volume 33, Issue 1, January–February 2015, The CRISPR/Cas9 system for plant genome editing and beyond. MIT TECHNOLOGY REVIEW 08/10/15 DuPont Predicts CRISPR Plants on Dinner Plates in Five Years. Powerful and possibly unregulated, gene editing starts new boom in GMOs.
Gene editing could increase the number and type of genetically modified crops. Agricultural biotech giants are starting to make moves into CRISPR gene editing, saying they’ll be selling seeds engineered with the technology by the end of this decade. DuPont said today it entered an agreement with Caribou Biosciences, a spin-off from the laboratory of Jennifer Doudna at the University of California, Berkeley, who carried out key work on CRISPR-Cas9, a technology that provides something like a find-and-replace feature for DNA. GENETIC LITERACY PROJECT 16/12/15 European researchers in limbo waiting for EU’s decision on whether CRISPR plants are GMOs. GENETIC LITERACY PROJECT 14/01/16 How should Europe regulate CRISPR, new breeding technologies (NBTs)? FASTCOEXIST 15/03/16 CRISPR Is Going To Revolutionize Our Food System—And Start A New War Over GMOs. TEST BIOTECH 22/01/15 Synthetic gene technologies and their application regarding plants and animals in agriculture. PEERJ 17/04/14 The evolutionary history and diagnostic utility of the CRISPR-Cas system within Salmonella enterica ssp. enterica.
NEWS MEDICAL 12/12/15 CRISPR-Cas9 can help knock out genes in exotic animals. The simplicity of CRISPR-Cas9 gene editing will soon make studying the genes of any organism, from the simplest slime mold to the octopus, as easy as it now is to study the genes controlling development in standard lab animals such as nematodes, fruit flies, frogs and mice. A new study from the University of California, Berkeley, illustrates the ease with which CRISPR-Cas9 can knock out genes in exotic animals - in this case, an amphipod or sandhopper - to learn how those genes control growth and development.
Researchers wanted to know which genes control the development of appendages on each segment of the amphipod, whose body is like a Swiss army knife with each segment bearing a different blade or tool as an appendage. In less time than it would have taken two years ago to knock out one gene in the animal, UC Berkeley researchers knocked out six, shedding light on the basic genetic mechanisms that determine leg anatomy in the evolution of animals.
Master regulatory genes Source: GREENPEACE 20/01/16 New GM food could end up on your plate untested and unlabelled. After two decades of commercial use, Europeans still can’t stomach genetically modified (GM) food. But their producers may have found a way to bypass public opposition and safety regulation. PLOS 15/07/15 A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. Abstract The number of fully sequenced fungal genomes is rapidly increasing. Since genetic tools are poorly developed for most filamentous fungi, it is currently difficult to employ genetic engineering for understanding the biology of these fungi and to fully exploit them industrially. For that reason there is a demand for developing versatile methods that can be used to genetically manipulate non-model filamentous fungi.
FAS USDA 07/12/15 Germany: Government Agencies Argue About Classification of CRISPR and Other NBT's. PNAS 25/08/15 Opinion: Is CRISPR-based gene drive a biocontrol silver bullet or global conservation threat? ENVIRONMENT AGENCY AUSTRIA - FEV 2014 - New plant breeding technique - Risks associated with their application. FOOD SAFETY MAGAZINE 21/04/16 Genetically Modified Mushrooms One Step Closer to Store Shelves. News | April 21, 2016 By Staff The U.S. Mol Plant Pathol. 2016 Jan 25. Development of broad virus resistance in non-transgenic cucumber using CRISPR/Cas9 technology.
FRONTIERS IN PLANT SCIENCE 12/04/16 CRISPR/Cas9: A Tool to Circumscribe Cotton Leaf Curl Disease. PLANT BIOTECHNOLOGY 05/09/16 An Agrobacterium-delivered CRISPR/Cas9 system for high-frequency targeted mutagenesis in maize. MOLECULAR PLANT PATHOLOGY 08/10/16 Engineering of CRISPR/Cas9-mediated potyvirus resistance in transgene-free Arabidopsis plants. BAY AREA LYME FOUNDATION 16/10/16 CRISPR Technology: A New Approach for Eradicating Borrelia. FAS USDA 30/10/15 CRISPR and other NBT’s classified as GMO‘s. Insect Sci. 2016 Jun;23(3):469-77. CRISPR/Cas9-mediated targeted gene mutagenesis in Spodoptera litura. FRONT. PLANT 01/06/16 An Overview of CRISPR-Based Tools and Their Improvements: New Opportunities in Understanding Plant–Pathogen Interactions for Better Crop Protection.