Feng Zhang及同事將可定制的TALE DNA結(jié)合域與光敏“隱花色素-2”蛋白及其來(lái)自擬南芥的相互作用伙伴CIB1結(jié)合在了一起，從而生成了一個(gè)光遺傳“雙雜合”系統(tǒng)（他們將其稱為L(zhǎng)ITEs,，即“光可誘導(dǎo)的轉(zhuǎn)錄效應(yīng)物”）,。LITEs不需要其他輔因子,，容易被定制來(lái)以很多位點(diǎn)為目標(biāo)，并且還能快速地,、可逆地被激活,。它們還可被打包到病毒載體內(nèi)，定向輸送到特定細(xì)胞類(lèi)群中,。作者將這一系統(tǒng)應(yīng)用到了小鼠的原代神經(jīng)元中和清醒小鼠的腦中,，來(lái)調(diào)制內(nèi)源基因表達(dá)和定位表觀染色質(zhì)修飾。這一LITE系統(tǒng)為內(nèi)源細(xì)胞過(guò)程的光遺傳控制建立了一個(gè)新穎模型,。（生物谷Bioon.com）
生物谷推薦英文摘要：
Nature doi: 10.1038/nature12466
Optical control of mammalian endogenous transcription and epigenetic states
Silvana Konermann, Mark D. Brigham,Alexandro E. Trevino,Patrick D. Hsu, Matthias Heidenreich,Le Cong,Randall J. Platt,David A. Scott,George M. Church & Feng Zhang
The dynamic nature of gene expression enables cellular programming, homeostasis and environmental adaptation in living systems. Dissection of causal gene functions in cellular and organismal processes therefore necessitates approaches that enable spatially and temporally precise modulation of gene expression. Recently, a variety of microbial and plant-derived light-sensitive proteins have been engineered as optogenetic actuators, enabling high-precision spatiotemporal control of many cellular functions. However, versatile and robust technologies that enable optical modulation of transcription in the mammalian endogenous genome remain elusive. Here we describe the development of light-inducible transcriptional effectors (LITEs), an optogenetic two-hybrid system integrating the customizable TALE DNA-binding domain with the light-sensitive cryptochrome 2 protein and its interacting partner CIB1 from Arabidopsis thaliana. LITEs do not require additional exogenous chemical cofactors, are easily customized to target many endogenous genomic loci, and can be activated within minutes with reversibility. LITEs can be packaged into viral vectors and genetically targeted to probe specific cell populations. We have applied this system in primary mouse neurons, as well as in the brain of freely behaving mice in vivo to mediate reversible modulation of mammalian endogenous gene expression as well as targeted epigenetic chromatin modifications. The LITE system establishes a novel mode of optogenetic control of endogenous cellular processes and enables direct testing of the causal roles of genetic and epigenetic regulation in normal biological processes and disease states.
