Identification of genes involved in cell invasion by using a library of randomized hybrid ribozymes

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Abstract
Libraries of RNA helicase-coupled randomized ribozymes are a powerful tool for the identification of functional genes. We have demonstrated the usefulness of this functional gene-discovery system by identifying genes involved in tumor invasion, a process that is an essential feature of tumor metastasis: the spread of cancer cells from the original tumor to other sites in the body that imposes serious problems in the prognosis and treatment of cancer. Using a filter-based invasion assay in vitro, we isolated ribozymes that enhanced the invasive properties of NIH 3T3 fibroblasts. Sequence analysis of selected clones and a database search revealed that genes such as the gene for Gem GTPase and uncharacterized genes that resemble genes for myosin phosphatase and protein-tyrosine-phosphatase are involved in cell invasion. Our system for gene identification by using ribozymes and the functional analysis of target genes should help to clarify the complex mechanisms of invasion and metastasis and might provide information that is relevant to cancer therapy.