RNA Interference Functional Oncogenomics Laboratory (RIFOL)
Functional OncoGenomics is the research area directed at elucidating the molecular causes of aberrant functions in cancer cells. This knowledge is of great importance for devising new, more effective treatments of cancer. The biological process of RNA interference (RNAi) provides a wealth of opportunities for Functional Oncogenomics research. Using RNAi, the genetic cause of a particular function can be assessed by individually suppressing gene products in cancer cells and studying changes in the behavior of these cancer cells. Pinpointing gene products that are functionally associated with processes in cancer cells contributes to a better understanding of cancer cell biology as well as identifying targets for therapy.
In october 2006, the VUmc Cancer Center Amsterdam (CCA) established a core facility for Functional OncoGenomics research by RNAi library screening. This RNA Interference Functional Oncogenomics Laboratory (RIFOL) is located in the laboratories of the section Oncogenomics of the department of Medical Oncology in the VUmc CCA research building. The RIFOL was set up with financial support from the Stichting VUmc CCA. RIFOL's mission is to provide researchers at the VUmc, in particular within CCA with facilities to conduct functional siRNA library screens.
The RIFOL participates in the Genome-Wide RNAi Global Initiative , an alliance of Thermo Fisher Scientific and leading international research centers pioneering the use of whole-genome RNAi screening. The mission of the RNAi Global Initiative is to combine the powerful new technology of genome-wide siRNA screening with international scientific exchange and the collaboration of leading research institutions in order to rapidly accelerate basic biological and medical discovery. The RNAi Global Initiative provides a forum for member institutions to share research protocols, establish experimental standards and develop mechanisms for exchanging and comparing screening data. Click here for the press release reporting on the VUmc-RIFOL's membership of the RNAi Global Initiative.
RNA Interference (RNAi) is a conserved biological process by which a double-stranded RNA molecule specifically suppresses production of a protein in a cell by targeting its encoding mRNA. This gene expression-silencing phenomenon was first observed in plants, although it was originally termed "co-suppression" and its mechanism was not recognized at that time. The first crucial breakthrough in understanding the RNAi mechanism came in 1998, when Andrew Fire and Craig Mello discovered that the RNAi pathway is triggered by double-stranded RNA. In 2006, Fire and Mello were awarded the Nobel Prize in Physiology or Medicine for this important discovery. In subsequent years, the RNAi process was further resolved. RNAi was found to be active in every cell of virtually every multi-cellular organism, including humans. Methods were developed to exploit the RNAi machinery in mammalian cells to specifically silence individual genes. This created a revolution in biology and medicine, opening up previously unexplored and even unsuspected areas of research. In 2002, Science magazine heralded RNAi technology the scientific "Breakthrough of the Year". In only a few more years, RNAi research evolved from the first biochemical understanding of the pathway to RNAi-based experimental medicines being tested in human clinical trials.
The RIFOL is the core facility for RNAi library screening in the VUmc Cancer Center Amsterdam. It serves primarily researchers of VUmc departments participating in the VUmc CCA. Other academic research groups can obtain access to the RIFOL facilities through collaboration with VUmc CCA departments. A VUmc CCA program advisory committee prioritizes proposals for new screening projects.
The RIFOL provides VUmc CCA researchers access to the Dharmacon siARRAY Human Genome Library. This library includes siRNA reagents that target over 21,000 unique genes in the human genome. Use of this library enables VUmc CCA researchers to perform genome-wide functional screens. The RIFOL works according to a partial cost-recovery model. Use of the laboratory facilities and support by RIFOL personnel is free for VUmc CCA users. Users are charged only for liquid handling consumable costs and library consumption. The RIFOL expects users to actively participate in the RIFOL team, exchanging protocols and experiences to maintain and improve the quality of our library screens.
The RIFOL personnel assist screening efforts by providing access to the libraries, by operating and maintaining equipment, by providing guidance in performing functional assays and by advising on data analysis. Users develop and perform their own assays with their own supplies. They are primarily responsible for their screens and report directly to their department. The RIFOL is equipped to support mammalian cell-based high-throughput screens, at maximum ML-2 biocontainment level.
Current RIFOL Projects
Chemo-radiation resistance in HNSCC (Sanne de Kemp; Dept. Otolaryngology)
Therapy resistance in invasive glioma (Petra van der Stoop; Dept. Neurosurgery)
Radiation resistance in prostate cancer (Jasmina Hodzic, Dept. Medical Oncology)
Cisplatin resistance in NSCLC (Ellen Siebring-van Olst; Dept. Pulmonary Diseases)
Doxorubicin resistance in osteosarcoma (Jantine PosthumaDeBoer, Dept. Orthopedic Surgery)
Radiation resistance in lung cancer (Remco Nagel, Dept. Otolaryngology)
Synthetic lethality sister chromatid cohesion defects (Job de Lange, Dept. Clinical Genetics).
Targeted therapy of precancerous fields in head and neck mucosa (Vicky de Boer, Dept. Otolaryngology)