CSIR-Indian Institute of Chemical Biology

Researchers have modified a gene editing tool to serve as a highly sensitive diagnostic test for the presence of the SARS-CoV-2 virus. An engineered CRISPR-based method that finds RNA from SARS-CoV-2, the virus that causes COVID-19, promises to make testing for that and other diseases fast and easy. Collaborators at Rice University and the University of Connecticut further engineered the RNA-editing CRISPR-Cas13 system to boost their power for detecting minute amounts of the SARS-CoV-2 virus in biological samples without the time-consuming RNA extraction and amplification step necessary in gold-standard PCR testing. The new platform was highly successful compared to PCR, finding 10 out of 11 positives and no false positives for the virus in tests on clinical samples directly from nasal swabs. The researchers showed their technique finds signs of SARS-CoV-2 in attomolar (10-18) concentrations. The study led by chemical and biomolecular engineer Xue Sherry Gao at Rice's George R. Brown School of Engineering and postdoctoral researchers Jie Yang of Rice and Yang Song of Connecticut appears in Nature Chemical Biology. Cas13, like its better-known cousin Cas9, is part of the system by which bacteria naturally defend themselves against invading phages. Since its discovery, CRISPR-Cas9 has been adapted by scientists to edit living DNA genomes and shows great promise to treat and even cure diseases. And it can be used in other ways. Cas13 on its own can be enhanced with guide RNA to find and snip target RNA sequences, but also to find "collateral," in this case the presence of viruses like SARS-CoV-2. "The engineered Cas13 protein in this work can be readily adapted to other previously established platforms," Gao said. "The stability and robustness of engineered Cas13 variants make them more suitable for point-of-care diagnostics in low-resource setting areas when expensive PCR machines are not available." Yang said wild-type Cas13, drawn from a bacterium, Leptotrichia wadei, cannot detect attomolar level of viral RNA within a time frame of 30 to 60 minutes, but the enhanced version created at Rice does the job in about half an hour and detects SARS-CoV-2 in much lower concentrations than the previous tests. She said the key is a well-hidden, flexible hairpin loop near Cas13's active site. "It's in the middle of the protein near the catalytic site that determines Cas13's activity," Yang said. "Since Cas13 is large and dynamic, it was challenging to find a site to insert another functional domain." The researchers fused seven different RNA binding domains to the loop, and two of the complexes were clearly superior. When they found their targets, the proteins would fluoresce, revealing the presence of the virus. "We could see the increased activity was five- or six-fold over wild-type Cas13," Yang said. "This number seems small, but it's quite astonishing with a single step of protein engineering. "But that was still not enough for detection, so we moved the whole assay from a fluorescence plate reader, which is quite large and not available in low-resource settings, to an electrochemical sensor, which has higher sensitivity and can be used for point-of-care diagnostics," she said. With the off-the-shelf sensor, Yang said the engineered protein was five orders of magnitude more sensitive in detecting the virus compared to the wild-type protein. The lab wants to adapt its technology to paper strips like those in home COVID-19 antibody tests, but with much higher sensitivity and accuracy. "We hope that will make testing more convenient and with lower cost for many targets," Gao said. The researchers are also investigating improved detection of the Zika, dengue and Ebola viruses and predictive biomarkers for cardiovascular disease. Their work could lead to rapid diagnosis of the severity of COVID-19. "Different viruses have different sequences," Yang said. "We can design guide RNA to target a specific sequence that we can then detect, which is the power of the CRISPR-Cas13 system." But because the project began just as the pandemic took hold, SARS-CoV-2 was a natural focus. "The technology is quite amenable to all the targets," she said. "This makes it a very good option to detect all kinds of mutations or different coronaviruses." "We are very excited about this work as a combinational effort of structure biology, protein engineering and biomedical device development," Gao added. "I greatly appreciate all the efforts from my lab members and collaborators." Co-authors of the paper are Rice postdoctoral researcher Xiangyu Deng, undergraduate Jeffrey Vanegas and graduate student Zheng You; graduate students Yuxuan Zhang and Zhengyan Weng of the University of Connecticut; microbiology supervisor Lori Avery and Kevin Dieckhaus, a professor of medicine, of UConn Health; Yi Zhang, an assistant professor of biomedical engineering at the University of Connecticut; and Yang Gao, an assistant professor of biosciences at Rice. Xue Sherry Gao is the Ted N. Law Assistant Professor of Chemical and Biomolecular Engineering at Rice. The National Science Foundation (2031242, 2103025), the Welch Foundation (C-1952, C-2033-20200401), and the Cancer Prevention and Research Institute of Texas (RR190046) supported the research.

Preclinical evaluation of HetSiFA®-based theranostic candidates to be initial focus THE WOODLANDS, Texas--(BUSINESS WIRE)-- Fuzionaire Diagnostics, Inc. ("Fuzionaire Dx") today announced a collaborative research agreement with McMaster University ("McMaster") under which theranostic radiopharmaceuticals developed with Fuzionaire Dx's HetSiFA® platform will be evaluated at McMaster's comprehensive preclinical nuclear medicine facilities. The collaboration expands Fuzionaire Dx’s capabilities to evaluate oncologic theranostics made from its HetSiFA platform. The collaboration will begin with the biological and preclinical evaluation of multiple theranostic candidates targeting an undisclosed type of cancer and selection of promising candidates for further development. Fuzionaire Dx creates theranostic candidates by combining its HetSiFAs, or heteroaromatic silicon-fluoride acceptors, with cancer-targeting peptides and other ligands. With the HetSiFA library, new theranostic candidates are created within weeks. The HetSiFA platform is compatible with fluorine-18, the leading radionuclide used in positron-emission-tomography (PET) imaging, and leading alpha- and beta-emitting radionuclides used for therapy, providing unique flexibility in creating new candidates. Theranostics is a term that describes a "see it, treat it" approach to cancer that uses one radioactive drug to reveal tumor locations and a second radioactive drug to deliver therapy that kills cancer cells. Primary leadership for the scientific collaboration includes Christopher Waldmann, PhD, Chief Scientific Officer of Fuzionaire Dx, and Saman Sadeghi, PhD, Associate Professor, Chemistry & Chemical Biology, and Principal Investigator, Radiochemistry & Radiopharmaceuticals, at McMaster University. About Fuzionaire Diagnostics Fuzionaire Diagnostics is a radiopharmaceutical company on a mission to develop accessible theranostics to fight cancer. With origins in Nobel laureate Robert Grubbs' lab at Caltech, our patented HetSiFA® library can create theranostic radiopharmaceutical candidates from disease-targeting peptides and other ligands within weeks. Our platform also offers unique flexibility in the choice of the radionuclide used for imaging or therapy as well as manufacturing advantages in a clinical setting that enable the wider use of these important precision treatments. For more, visit us at: . About McMaster University McMaster University is among Canada’s most research-intensive universities and is consistently ranked as one of the world’s Top 100 universities. Together, our researchers, students and staff advance human and societal health and well-being, creating a Brighter World.

Ottawa, Canada, April 13, 2021 (GLOBE NEWSWIRE) -- Orion Biotechnology Canada Ltd., a clinical stage biotechnology company focused on precision engineering small protein therapeutics, today announced the addition of four experts to further strengthen its Scientific Advisory Board (SAB). These new SAB members include Dr. Christian Heinis, Dr. Laurent Jespers, Dr. Sebastian Stintzing and Dr. Josep Tabernero. The Orion Biotechnology SAB is comprised of leading experts in cell receptor pharmacology, biochemistry, translational medicine and research and development (R&D) in oncology and immunology.   These individuals serve as key resources for Orion Biotechnology, providing insights as well as offering advice on the company’s drug discovery strategy and technology. “We are honoured to welcome Drs. Heinis, Jespers, Stintzing and Tabernero as members of our SAB.  Their input will prove invaluable as we begin the next phase of our development” said Dr. Ian McGowan - Chief Medical Officer. “Each member’s expertise will expand our knowledge base and further inform our research and clinical development activities, going forward.” added Dr. Oliver Hartley – VP, Drug Discovery. Orion Biotechnology is pioneering a new analog-based approach to targeting high-value G-Protein Coupled Receptors (GPCRs) that utilizes native peptide ligands as a scaffold on which to precision engineer drug leads with enhanced potency and tailored signalling activity.  This approach uniquely positions Orion to focus on complex GPCRs which have not been successfully drugged by standard discovery approaches, including small molecules and monoclonal antibodies. About Orion Biotechnology’s New Scientific Advisory Board (SAB) Members Dr. Christian Heinis is a Professor in Bioorganic Chemistry at the Ecole Polytechnique Fédérale de Lausanne (EPFL) in Switzerland. His laboratory is developing combinatorial methods for creating therapeutics based on cyclic peptides and macrocyclic compounds. He has over 15 years of experience in peptide engineering, in vitro evolution, and drug development. Dr. Heinis is a co-founder of Bicycle Therapeutics, and co-director of the NCCR Chemical Biology, a research network of around twenty groups in the Lake Geneva area. Dr. Heinis was trained in chemistry and biochemistry at the ETH Zurich. He did his PhD with Prof. Dario Neri (ETH Zurich), a first post-doc with Prof. Kai Johnsson (EPFL) and a second post-doc with Sir Greg Winter (LMB-MRC in Cambridge, UK). Dr. Josep Tabernero is Head of the Medical Oncology Deparment at the Vall d’Hebron University Hospital, Director of the Vall d’Hebron Institute of Oncology (VHIO), and Professor of Medicine at UVic-UCC.  He has been a Principal Investigator of several Phase 1 pharmacodynamic studies and translational projects with tumor-directed targeted and immune-based therapies.  Dr. Tabernero serves on the Editorial Boards of various top tier journals including Annals of Oncology, ESMO Open, Cancer Discovery, Clinical Cancer Research, Cancer Treatment Reviews, and Nature Reviews Clinical Oncology. Dr. Tabernero is currently the Past President of the European Society for Medical Oncology´s (ESMO), and also serves on its Public Policy as well as Cancer Medicines Committees.  He is also a member of the American Association for Cancer Research (AACR) and the American Society of Clinical Oncology (ASCO). Dr. Laurent Jespers  serves as CSO for an allogeneic cell therapy biotech start-up based in Cambridge, UK. During his 12 year career at GSK he led research teams as VP of Biopharm Innovation, and as VP of Discovery in Cell & Gene Therapy. Dr. Jespers drove the design and therapeutic application of over 10 therapeutic modalities (antibody, peptide, gene therapies, and autologous cell therapies) from concept to preclinical development in diseases areas such as Oncology Immuno-Inflammation, Metabolic, and Respiratory. Prior to GSK, he was Director of Discovery Engineering at Domantis Ltd, a company developing human antibody variable fragments. Dr. Jespers earned his PhD in biochemistry from the University of Louvain, Belgium, and completed his post-doctoral training in protein engineering at the MRC-Laboratory of Molecular Biology, Cambridge, UK.  Dr. Jespers co-developed the guided antibody humanisation approach which contributed to the development of Humira, the first fully human mAb to be approved by the U.S. Food and Drug Administration. Dr. Sebastian Stintzing  is Professor of Medicine and Head of the Department of Hematology, Oncology, and Tumor Immunology (CCM) of Charité Universitaetsmedizin Berlin. His research focuses on predictive and prognostic biomarkers in the treatment of gastrointestinal cancer. He has coordinated the translational science and clinical course of several studies. In 2012, Dr. Stintzing received the prestigious Research Fellowship Award from the “Deutsche Krebshilfe” (German Cancer Aid) and in 2014, he received the scientific award of the AIO (working group of medical oncology within the German Cancer Society). Dr. Stintzing received his medical degree from the Friedrich-Alexander University of Erlangen-Nuremberg and earned his Postdoctoral Lecture Qualification (Habilitation) with a thesis on “Prognostic and predictive factors in the treatment of metastatic colorectal cancer “. Dr. Stintzing is member of several cancer associations (ESMO, ASCO, and DGHO) as well as a member of the S3-guideline committee for colorectal cancer in Germany. About Orion Biotechnology Holding SA  Orion Biotechnology Canada Ltd, is a clinical stage biopharmaceutical company focused on the discovery and development of precision engineered small protein therapeutics – a promising new class of drugs designed to modulate immune response and deliver the next generation of immunotherapy treatment for cancer and other serious diseases. Leveraging its proprietary drug discovery platform to facilitate rapid and low-cost development of its molecules, Orion is pursuing a range of novel targets including chemokine and other G-Protein Coupled Receptors (GPCR’s). Orion’s lead candidate, OB-002, is a first-in-class chemokine analog and best in class CCR5 antagonist. Our close ties to diverse institutions and experts around the globe, as well as our proprietary drug discovery platform, continue to stimulate the rapid discovery of promising new treatments.  Learn more at .