Optogenetic Inhibitor of the Transcription Factor CREB.
Optogenetic Inhibitor of the Transcription Factor CREB.
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DOI：
10.1016/j.chembiol.2015.09.018
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发表时间：
2015-11-19
影响因子：
--
通讯作者：
Woolley GA
中科院分区：
文献类型：
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作者：
Ali AM;Reis JM;Xia Y;Rashid AJ;Mercaldo V;Walters BJ;Brechun KE;Borisenko V;Josselyn SA;Karanicolas J;Woolley GA
目前的光遗传学转录控制方法不能模拟内源性转录因子的活性，内源性转录因子在基因组的许多位点与其他因素相互作用。光遗传学控制显性内源转录因子的负性版本提供了一种模拟基因表达自然调控的机制。在这里，我们描述了opto-DN-CREB，一种蓝光控制的转录因子CREB抑制剂，通过将显性负抑制剂a -CREB与光活性黄蛋白（PYP）融合而产生。PYP中光驱动的构象变化可以防止A-CREB和CREB之间形成线圈，从而激活CREB。在体外、HEK293T细胞和神经元中，蓝光能够控制CREB靶点NR4A2和c-Fos的表达，从而表征了CREB功能的光遗传学控制。显性负抑制剂存在于许多转录因子；将这些与光遗传结构域联系起来，为自然转录事件的时空控制提供了一种通用方法。
Current approaches for optogenetic control of transcription do not mimic the activity of endogenous transcription factors, which act at numerous sites in the genome in a complex interplay with other factors. Optogenetic control of dominant negative versions of endogenous transcription factors provides a mechanism for mimicking the natural regulation of gene expression. Here we describe opto-DN-CREB, a blue light controlled inhibitor of the transcription factor CREB created by fusing the dominant negative inhibitor A-CREB to photoactive yellow protein (PYP). A light driven conformational change in PYP prevents coiled-coil formation between A-CREB and CREB, thereby activating CREB. Optogenetic control of CREB function was characterized in vitro, in HEK293T cells, and in neurons where blue light enabled control of expression of the CREB targets NR4A2 and c-Fos. Dominant negative inhibitors exist for numerous transcription factors; linking these to optogenetic domains offers a general approach for spatiotemporal control of native transcriptional events.