A Conformational Restriction Strategy for the Control of CRISPR/Cas Gene Editing with Photoactivatable Guide RNAs.

The CRISPR/Cas system is one of the most powerful tools for gene editing. However, approaches for precise control of genome editing and regulatory events are still desirable. Here, we report the spatiotemporal and efficient control of CRISPR/Cas9- and Cas12a-mediated editing with conformationally restricted guide RNAs (gRNAs). This approach relied on only two or three pre-installed photo-labile substituents followed by an intramolecular cyclization, representing a robust synthetic method in comparison to the heavily modified linear gRNAs that often require extensive screening and time-consuming optimization. This tactic could direct the precise cleavage of the genes encoding green fluorescent protein (GFP) and the vascular endothelial growth factor A (VEGFA) protein within a predefined cutting region without notable editing leakage in live cells. We also achieved light-mediated myostatin (MSTN) gene editing in embryos, wherein a new bow-knot-type gRNA was constructed with excellent OFF/ON switch efficiency. Overall, our work provides a significant new strategy in CRISPR/Cas editing with modified circular gRNAs to precisely manipulate where and when genes are edited.

Highly selective colorimetric and electrochemical Pb2+ detection based on TTF-pi-pyridine derivatives.

[reaction: see text] The pyridineethenyl-substituted tetrathiafulvalene (TTF) compounds, 4-(4-pyridineethenyl)tetrathiafulvalene (1a) and 4,4'(5')-[bis-(4-pyridineethenyl)]tetrathiafulvalene (2a) together with the styryl-substituted TTF compounds, 4-styryltetrathiafulvalene (1b) and 4,4'(5')-bis-styryltetrathiafulvalene (2b), have been designed and synthesized. All these compounds exhibit strong absorption bands in the range of 370 to 550 nm, which are assigned to the intramolecular charge-transfer transition from the HOMO in TTF to the LUMO in the pyridyl or phenyl group. Compared to compounds 1b and 2b, the pyridineethenyl-substituted TTF compounds 1a and 2a show remarkable sensing and coordinating properties to Pb2+. With the addition of micromolar concentrations of Pb2+ to the solution, 1a or 2a displays dramatic changes in the UV-vis absorption spectrum, 1H NMR spectrum, and redox property.