NEW YORK (Reuters Health) - Although gene silencing by RNA interference (RNAi) is a potentially valuable means of genetic analysis, technological issues have limited its large-scale use in mammalian cells. Now, two research groups seem to have overcome this problem by creating tools that facilitate RNAi mass-screening of mammalian genes.
RNAi has been used to turn off virtually every gene in two popular lab organisms -- nematode worms and fruitflies -- Dr. Andrew Fraser, from The Wellcome Trust Sanger Institute in Hinxton, UK, notes in an editorial accompanying the reports in the March 25th issue of Nature. “Such genome-wide RNAi surveys of gene function have remained out of reach in mammals -- until now.”
Research teams led by Dr. Patrick J. Paddison, from Cold Spring Harbor Laboratory in New York, and by Dr. Katrien Berns, from The Netherlands Cancer Institute in Amsterdam, created retrovirus-based RNAi libraries that covered about a third of human genes. The specific genes were selected based on their potential to cause disease.
Dr. Paddison’s team generated a library that targeted 9610 human and 5563 mouse genes. A genetic screen designed to report defects in human proteasome function was used to validate the library.
“Our results suggest that our large-scale RNAi library can be used in specific, genetic applications in mammals, and will become a valuable resource for gene analysis and discovery,” the investigators note.
Dr. Berns’ team focused on genes that affected the function of p53, the well-known tumor suppressor gene. One known and five new modulators of p53-dependent proliferation arrest were identified with the RNAi screening technique.
“We will no doubt see an explosion in RNAi screening of mammalian cells over the coming months,” Dr. Fraser states. “As with any genetic screen, the power of each RNAi screen depends on the appropriate choice of functional readout, and that will require development of a variety of cell-based assays.”
Source: Nature 2004;428:375-378,427-437. [ Google search on this article ]
MeSH Headings:Biological Phenomena: Biological Phenomena, Cell Phenomena, and Immunity: Community Health Services: Data Collection: Diagnosis: Diagnostic Services: Environment and Public Health: Epidemiologic Methods: Health Care Facilities, Manpower, and Services: Genetic Screening: Health: Health Occupations: Health Services: Health Services Administration: Health Surveys: Information Science: Mass Screening: Medicine: Microbiologic Phenomena: Investigative Techniques: Population Characteristics: Preventive Health Services: Preventive Medicine: Public Health: Quality of Health Care: Specialties, Medical: Viral Interference: Public Health Practice: Health Care Quality, Access, and Evaluation: Health Care Evaluation Mechanisms: Viral Physiology: Diagnostic Techniques and Procedures: Analytical, Diagnostic and Therapeutic Techniques and Equipment: Biological Sciences: Health Care: Information ScienceCopyright © 2002 Reuters Limited. All rights reserved. Republication or redistribution of Reuters content, including by framing or similar means, is expressly prohibited without the prior written consent of Reuters. Reuters shall not be liable for any errors or delays in the content, or for any actions taken in reliance thereon. Reuters and the Reuters sphere logo are registered trademarks and trademarks of the Reuters group of companies around the world.