Optogenetic Control of Gene Expression Using Cryptochrome 2 and a Light-Activated Degron.
Methods in molecular biology (Clifton, N.J.)(2020)
摘要
Optogenetic tools allow for use of light as an external input to control cellular processes. When applied to regulate the function of transcription factors, optogenetic approaches provide a tunable, reversible, and bidirectional method to control gene expression. Herein, we present a detailed method to induce gene expression in mammalian cells using the light dependent dimerization of cryptochrome 2 (CRY2) and CIB1 to complement a split transcription factor. We also describe a protocol to disrupt gene expression with light by fusing a dimeric transcription factor to CRY2. When combined with a light-induced degron attached to the gene product, this method allows for rapid modulation of target protein abundance.
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关键词
CRY2,Degron,Light,Optogenetics,Transcription
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