Optical Control of Mammalian Endogenous Transcription and Epigenetic States

By Silvana Konermann, Mark D. Brigham, Alexandro E. Trevino, P. Hsu, Matthias Heidenreich, Le Cong, Randall J. Platt, David A. Scott, George M. Church, Feng Zhang. Nature, 2013

830 citations recorded by the source corpus.

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.

Subfields: Biotechnology, Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/65367984769c4556a05f5d91fb84648290aa3748

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Citations in the corpus

Transcriptional enhancers: from properties to genome-wide predictions (2014, background): “Through these domains, transcriptional activators (part Aa ), repressive transcription factors (TFs) 117,137,139 (part Ab ) or chromatin-modifying domains 138,139 (part Ac ) can be recruited to genomic regulatory regions to modulate the transcription of target genes.”
Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA (2014, background): “Moreover, a catalytically inactive Cas9 can serve as an RNA-guided DNA-binding platform to target effector domains and modulate endogenous transcription (Gilbert et al., 2013; Konermann et al., 2013; Maeder et al., 2013; Perez-Pinera et al., 2013; Qi et al., 2013).”
CRISPR-Cas9 Knockin Mice for Genome Editing and Cancer Modeling (2014, methodology): “…end-joining (NHEJ), resulting in insertions and deletions (indels) (Bibikova et al., 2002), or ho- mology-directed repair (HDR) (Bibikova et al., 2003; Bibikova et al., 2001; Jasin et al., 1985; Rudin et al., 1989), resulting in precise sequence substitution in the presence of a repair…”
Development and Applications of CRISPR-Cas9 for Genome Engineering (2014, positive): “Fusing them to modular, sequence-agnostic functional effector domains allows flexible recruitment of desired perturbations, such as transcriptional activation, to a locus of interest (Xu and Bestor, 1997; Beerli et al., 2000a; Konermann et al., 2013; Maeder et al., 2013a; Mendenhall et al., 2013).”
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