Chemical control over membrane-initiated steroid signaling with a DNA nanocapsule

Veetil, Aneesh Tazhe; Jani, Maulik S; Krishnan, Yamuna

doi:10.1073/pnas.1712792115

Cited by 12 publications

(9 citation statements)

References 27 publications

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“…Using a well-characterized, icosahedral DNA nanocapsule reporter system (SI Fig. 3 & 4) 28,29 , we found that uncatalyzed thioldisulfide exchange mediated by small biologically available thiols was negligible.…”

Section: Tdx Detects Disulfide Exchange In Late Endosome In Vivomentioning

confidence: 97%

DNA nanodevices map enzymatic activity in organelles

et al. 2019

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Cellular reporters of enzyme activity are based on either fluorescent proteins or small molecules. Such reporters provide information corresponding to wherever inside cells the enzyme is maximally active and preclude minor populations present in sub-cellular compartments. Here we describe a chemical imaging strategy to selectively interrogate minor, sub-cellular pools of enzymatic activity. This new technology confines the detection chemistry to a designated organelle, enabling imaging of enzymatic cleavage exclusively within the organelle. We have thus quantitatively mapped disulfide reduction exclusively in endosomes in C. elegans and identified that exchange is mediated by minor populations of enzymes PDI-3 and TRX-1 resident in endosomes. Impeding intra-endosomal disulfide reduction by knocking down TRX-1 protects nematodes from infection by Corynebacterium diphtheriae, revealing the importance of this minor pool of endosomal TRX-1. TRX-1 also mediates endosomal disulfide reduction in human cells. A range of enzymatic cleavage reactions in organelles are amenable to analysis by this new reporter strategy.Minor populations of proteins and protein complexes perform critical functions for the cell. For example, a minor population of the epidermal growth factor receptor present on exosomes mediates intercellular communication 1 . A small fraction of mammalian target of rapamycin, present on lysosomes is responsible for nutrient sensing by the cell 2 . A minor population of the KDEL receptor present in the Golgi apparatus performs the critical function of retrieving ER-resident proteins from the Golgi apparatus 3 . Small molecules that function as fluorescent reporters of enzymatic activity use highly specific and rapid Reprints and permission information is available online at www.nature.com/reprints.

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Section: Tdx Detects Disulfide Exchange In Late Endosome In Vivomentioning

confidence: 97%

DNA nanodevices map enzymatic activity in organelles

et al. 2019

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“…Immunostaining was done as described previously 58 . Briefly, Human monocyte derived macrophages (HMDMs) were differentiated on a glass bottom 3.5 cm imaging dish and treated with or without 100 ng/mL LPS for 24 hours.…”

Section: Immunostaining Of Myeloperoxidase (Mpo)mentioning

confidence: 99%

A DNA-based fluorescent reporter maps HOCl production in the maturing phagosome

et al. 2018

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Phagocytes destroy pathogens by trapping them in a transient organelle called the phagosome where they are bombarded with reactive oxygen (ROS) and reactive nitrogen species (RNS). Imaging reactive species within the phagosome would directly reveal the chemical dynamics underlying pathogen destruction. Here we introduce a fluorescent, DNA-based combination reporter, cHOClate which simultaneously images HOCl and pH quantitatively. Using cHOClate targeted to phagosomes in live cells we successfully map phagosomal production of a specific ROS i.e. hypochlorous acid (HOCl), as a function of phagosome maturation. We found that phagosomal acidification was gradual in macrophages and upon completion, HOCl was released in a burst. This revealed that phagosome-lysosome fusion was essential not only for phagosome acidification, but also to provide the chloride necessary for myeloperoxidase activity. This method can be expanded to image several kinds of ROS and RNS and be readily applied to identify how resistant pathogens evade phagosomal killing.

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“…Other than the simple i-motif structure, a DNA icosahedra was also designed and used for cargo-loading, which was further employed in cell imaging. − For example, they introduced three different endocytic ligand (folic acid (FA), galectin-3 (Gal3), and the Shiga toxin B-subunit (STxB)) to the quantum dot (QD)-loaded DNA icosahedra respectively through site-specific modification and demonstrated the feasibility to characterize the dynamics of endocytic pathways facilitated by Gal3 and STxB . Hence, the QD-loaded DNA icosahedra exhibited an excellent platform in real-time endocytic pathway mapping at single-particle level.…”

Section: Robots For Nanomedicinementioning

confidence: 99%

Robotic DNA Nanostructures

et al. 2020

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Over the past decade, DNA nanotechnology has spawned a broad variety of functional nanostructures tailored toward the enabled state at which applications are coming increasingly in view. One of the branches of these applications is in synthetic biology, where the intrinsic programmability of the DNA nanostructures may pave the way for smart task-specific molecular robotics. In brief, the synthesis of the user-defined artificial DNA nano-objects is based on employing DNA molecules with custom lengths and sequences as building materials that predictably assemble together by obeying Watson–Crick base pairing rules. The general workflow of creating DNA nanoshapes is getting more and more straightforward, and some objects can be designed automatically from the top down. The versatile DNA nano-objects can serve as synthetic tools at the interface with biology, for example, in therapeutics and diagnostics as dynamic logic-gated nanopills, light-, pH-, and thermally driven devices. Such diverse apparatuses can also serve as optical polarizers, sensors and capsules, autonomous cargo-sorting robots, rotary machines, precision measurement tools, as well as electric and magnetic-field directed robotic arms. In this review, we summarize the recent progress in robotic DNA nanostructures, mechanics, and their various implementations.

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Chemical control over membrane-initiated steroid signaling with a DNA nanocapsule

Cited by 12 publications

References 27 publications

DNA nanodevices map enzymatic activity in organelles

DNA nanodevices map enzymatic activity in organelles

A DNA-based fluorescent reporter maps HOCl production in the maturing phagosome

Robotic DNA Nanostructures

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