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Transcription errors in RNA, though rare and transient, lead to synthesis of defective proteins. Errors in RNA transcription can cause many human diseases, including those that are age-related.
Researchers at the National Cancer Institute and Center for Cancer Research, RNA Biology Laboratory, have developed an alternative to the standard Ames test (recommended by the FDA to assess the in vivo mutagenic and carcinogenic potential of a drug for drug approval). The new assay can detect the transient errors in transcription with the potential to test the level of toxicity caused by therapeutic compounds and prevent excessively toxic drugs from entering the market.
The assay is based on suppression of a mutation in the Cre recombinase, coupled with a Cre genetic reporter to monitor the activity of the recombinase. The Cre mutation results in an inactive enzyme; errors in RNA transcription of the Cre mutation restores the activity of the enzyme. Additionally, the genetic reporter of Cre activity leads to expression of a yellow fluorescent protein (YFP) and the mouse cell line in the invention assay has both the cre mutation source and cre activatable YFP gene. Activation of the YFP gene allows for the detection of transcription errors following exposure of the cells to compounds or other treatments. Further, exposure of the cells to elevated levels of manganese results in an increase of YFP expressing cells; these results are consistent with in vitro experiments demonstrating manganese can reduce transcription accuracy.
Researchers at the NCI seek licensing partners for the assay which detects mutagenicity in RNA synthesis caused by carcinogens or therapeutic candidates.
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