Photo-controllable multiple and orthogonal regulation of gene expression by chemically modified oligonucleotides

Abstract

Gene expression must be precisely regulated in cells and functional nucleic acids are the most widely utilized tools for gene manipulation. Photocontrol of how these nucleic acid tools work in the cellular environment can precisely manipulate gene expression through a non-invasive way. Here we report a methodology on multiplex photocontrol of functional nucleic acids to achieve totally temporal and orthogonal regulation of gene expression in living cells. We select two functional nucleic acid systems as examples, DNAzyme and CRISPR/Cas9, and demonstrate the power of light control for precise gene manipulation by rational design of chemically modified oligonucleotides through introduction of two photocleavable linkers. Unlike the previous modulation of functional nucleic acids by simply activating or deactivating, we successfully achieve versatile controlling patterns using light as the governing factor. This design represents a generalized pathway towards the photo-controllable functional nucleic acids, which greatly enriches the toolbox for optogenetic studies.