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New RNAi tools enable systematic studies of gene function

s tailored to match a fragment of a gene's unique DNA. This RNA can then bind to its gene target, rendering it inactive. In order to get the small RNAs into cells, TRC scientists packaged them in lentiviruses. "Across the spectrum of biomedicine, there is a need for tools that can be applied to diverse cell types. This is particularly true in cancer research," said Bill Hahn, assistant professor at Dana-Farber Cancer Institute and Harvard Medical School, associate member of the Broad Institute and a senior author of the study. "For TRC's library, lentiviral delivery is an especially effective means to meet this need."

The parallel analysis of thousands of genes using RNAi allows researchers to more readily pinpoint the genes that control a biological process. Therefore, TRC developed the high-throughput techniques and quality-control measures required for such genome-scale studies. "It is a distinct challenge to achieve consistent and cost-effective RNAi methods and we placed a strong emphasis on this part of the process," said David Sabatini, member of Whitehead Institute for Biomedical Research, assistant professor at Massachusetts Institute of Technology, associate member of the Broad Institute and a senior author. "In the quest to develop comprehensive tools for gene discovery in mice and humans, this technology will be a key piece in the puzzle."

To evaluate the RNAi library's performance, the scientists sampled a subset that targets approximately 1,000 human genes. They systematically inactivated these genes in a human cancer cell line to identify ones that regulate cell division during malignancy. Automated cellular imaging was used to efficiently identify dividing cells in thousands of samples. This approach uncovered more than 100 previously unknown growth regulators in addition to several known players, confirming the library's sensitivity as a vehicle for gene discovery.

"This critical new tool illustrates the requirement for acad
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Source:Broad Institute of MIT and Harvard


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