Julien Hiblot

Fast calcium-dependent fluorescent labeling for recording of neuronal activation

Nicola Porzberg, Jennifer Heck, Jonas Wilhelm, Joergen Benjaminsen et autres

Abstract Calcium transients encode cellular and neuronal activity across timescales ranging from milliseconds to hours, yet linking these transient signals to downstream molecular states remains a major challenge. We recently introduced Caprola, a calcium-dependent protein labeling tool that converts calcium transients into …

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Fast and Luminous: CLIP-tag2

Veselin Nasufović, Anamarija Pišpek, Stefanie Kühn, Manuel Bibrowski et autres

Abstract CLIP-tag is a self-labeling protein tag used for the specific fluorescence labeling of proteins. However, its low labeling speed and the poor cell permeability of its substrates result in low labeling efficiencies in live-cell applications. Here, we introduce a substrate optimized …

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Fast, Bright, and Reversible Fluorescent Labeling of Rhodamine-Binding Proteins

Julian Kompa, L Dornfeld, Nicola Porzberg, Soohyen Jang et autres

High Resolution Image Download MS PowerPoint Slide Rhodamine dyes conjugated to targeting ligands can yield exceptionally bright fluorescent probes for live-cell imaging. However, the limited permeability of such rhodamine derivatives restricts their broader applications, particularly in vivo . Here, we present Rho-tag …

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Glutamate indicators with increased sensitivity and tailored deactivation rates

Abhi Aggarwal, Adrian Negrean, Yang Chen, Rishyashring R. Iyer et autres

Understanding how neurons integrate signals from thousands of input synapses requires methods to monitor neurotransmission across many sites simultaneously. The fluorescent protein glutamate indicator iGluSnFR enables visualization of synaptic signaling, but the sensitivity, scale and speed of such measurements are limited by …

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Fast, Bright and Reversible Rhodamine Tags for Live-Cell Imaging

Julian Kompa, L Dornfeld, Nicola Porzberg, Soohyen Jang et autres

Abstract We present Rho-tag and SiR-tag, engineered protein tags derived from bacterial multidrug-resistance proteins that bind unsubstituted (silicon-) rhodamines with nanomolar affinity, enabling fast, reversible, and fluorogenic protein labeling. In live cells, Rho-tag labeling occurs within seconds — faster than HaloTag7 — …

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SNAP-tag2 for faster and brighter protein labeling

Stefanie Kühn, Veselin Nasufović, Jonas Wilhelm, Julian Kompa et autres

Abstract SNAP-tag is a powerful tool for labeling proteins with synthetic fluorophores in bioimaging. However, its utility in live-cell applications can be constrained by its relatively slow labeling kinetics and the limited cell permeability of its substrates. Here, we introduce improved labeling …

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A high-affinity split-HaloTag for live-cell protein labeling

Yin‐Hsi Lin, Julian Kompa, De‐en Sun, Runyu Mao et autres

Abstract We introduce a high-affinity split-HaloTag comprised of a short peptide tag (Hpep, 14 residues) and a large, inactive fragment (cpHaloΔ3). Hpep binds to cpHaloΔ3 spontaneously with nanomolar affinity, enabling subsequent labeling with fluorescent HaloTag ligands. The small size of Hpep facilitates …

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Improving split‐ HaloTag through computational protein engineering

Jonas Wilhelm, Lennart Nickel, Yin‐Hsi Lin, Julien Hiblot et autres

Split-HaloTag can be used to transform transient molecular interactions into permanent marks through chemical labeling, thereby enabling the recording of transient physiological events in individual cells. However, applications of split-HaloTag-based recorders can be limited by slow labeling rates. To address this issue, …

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Glutamate indicators with increased sensitivity and tailored deactivation rates

Kaspar Podgorski, Abhi Aggarwal, Adrian Negrean, Yang Chen et autres

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Glutamate indicators with increased sensitivity and tailored deactivation rates

Abhi Aggarwal, Adrian Negrean, Yang Chen, Rishyashring R. Iyer et autres

Abstract Identifying the input-output operations of neurons requires measurements of synaptic transmission simultaneously at many of a neuron’s thousands of inputs in the intact brain. To facilitate this goal, we engineered and screened 3365 variants of the fluorescent protein glutamate indicator iGluSnFR3 …

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Improving Split-HaloTag through Computational Protein Engineering

Jonas Wilhelm, Lennart Nickel, Yin‐Hsi Lin, Julien Hiblot et autres

Abstract Split-HaloTag can be used to transform transient molecular interactions into permanent marks through chemical labeling, thereby enabling the recording of transient physiological events in individual cells. However, applications of split-HaloTag-based recorders can be limited by slow labeling rates. To address this …

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SNAP-tag2: faster and brighter protein labeling

Stefanie Kühn, Veselin Nasufović, Jonas Wilhelm, Julian Kompa et autres

Abstract SNAP-tag is a powerful tool for labeling proteins with synthetic fluorophores in bioimaging. However, its utility in live-cell applications can be constrained by its relatively slow labeling kinetics and the limited cell permeability of its substrates. Here we introduce new labeling …

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