Organelle‐Specific Photoactivation of DNA Nanosensors for Precise Profiling of Subcellular Enzymatic Activity

Abstract

Understanding of the functions of enzymes in diverse cellular processes is important, but the design of sensors with controllable localization for in situ imaging of subcellular levels of enzymatic activity is particularly challenging. We introduce herein as patiotemporally controlled sensor technology that permits in situ localization and photoactivated imaging of human apurinic/apyrimidinic endonuclease 1 (APE1) within an intracellular organelle of choice (e.g., mitochondria or nucleus). The hybrid sensor platform is constructed by photoactivatable engineering of aD NA-based fluorescent probe and further combination with an upconversion nanoparticle and as pecific organelle localization signal. Controlled localization and NIR-light-mediated photoactivation of the sensor "on demand" effectively constrains the imaging signal to the organelle of interest, with improved subcellular resolution. We further demonstrate the application of the nanosensors for the imaging of subcellular APE1 translocation in response to oxidative stress in live cells.

Efficient delivery of a DNA aptamer-based biosensor into plant cells for glucose sensing through thiol-mediated uptake

“…Compared to natural receptors such as antibodies, aptamers are much smaller, more stable, and easier for scale-up manufacturing ( 19 , 20 ). The facile conjugation of the DNA with organelle-directing ( 21 ) or optical-responsive ( 22 ) groups has allowed spatiotemporal controls of imaging targets in different organelles using light. By virtue of these advantages, aptamer biosensors have been applied to environmental monitoring ( 23 ), food safety ( 24 ), medical diagnostics ( 25 ), and imaging ( 1 ).…”

Spatiotemporally Selective Molecular Imaging via Upconversion Luminescence‐Controlled, DNA‐Based Biosensor Technology

“…In addition, the dynamics of APE1 in subcellular localizations is correlated with tumor progression, metastasis, and therapeutic outcomes [51, 52] . Given the critical function of APE1, we validated the feasibility of our method for in situ APE1 imaging in an organelle of choice (Figure 6a) [53] . The sensing device contains a UV light‐activatable APE1‐sensing module (AP‐P), a UCNP as light transducer, and an organelle‐targeting ligand.…”

Controllable mitochondrial aggregation and fusion by a programmable DNA binder

“…1a). Here, the TPP cation is used to target mitochondrial and induce mito-fusion 24 . TPP-LDNA was synthesized based on a copper-catalyzed azide/alkyne cycloaddition reaction 25 between an azide-modi ed linkage DNA and an alkynyl-functionalized TPP cation (Supplementary Fig.…”