The ability to knockout genes in mice is a critical step in genetic research and therapeutics. "These negative features make it difficult to control precisely the expression level of a protein of interest in living cells and apply this method to mice." "The original AID system has two major drawbacks: leaky degradation and the requirement for a high dose of auxin," Kanemaki said. The AID system allows for a general, more efficient approach by which the target protein can be depleted in less than a few hours. In previous conditional gene knockout and siRNA studies, according to Kanemaki, it typically takes two or three days for a target protein to deplete. To initiate the degradation process, the researchers administered auxin, a plant hormone that helps regulate plant growth. Kanemaki and his team had previously developed an approach called the AID system, which uses a small protein tag, known as a degron, fused to proteins to induce degradation. "However, these technologies are not ideal for studying highly dynamic processes, such as cell cycle, differentiation, or neural activity, because of the slow rate of depletion of the protein of interest." Kanemaki, professor at the National Institute of Genetics in the Research Organization of Information and Systems (ROIS). "Conditional gene knockout and small interfering RNA (siRNA), which is used to silence proteins without knocking them out completely, has been employed in many studies," said Masato T. The Japan-based team published their results on November 11th in Nature Communications. Researchers can now more accurately and precisely target specific proteins in yeast, mammalian cells and mice to study how knocking down specific protein traits can influence physical manifestation in a cell or organism. The target protein is typically depleted in less than a few hours after treatment. Image: A degron-fused protein of interest is recognized for rapid degradation by a TIR1 mutant only in the presence of an auxin analog, which initiates the degradation process.
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