Optimization strategies and advances in the research and development of AAV‐based gene therapy to deliver large transgenes

Clinical and Translational Medicine, Год журнала: 2024, Номер 14(3)

Опубликована: Март 1, 2024

Adeno-associated virus (AAV)-based therapies are recognized as one of the most potent next-generation treatments for inherited and genetic diseases. However, several biological technological aspects AAV vectors remain a critical issue their widespread clinical application. Among them, limited capacity genome significantly hinders development AAV-based gene therapy. In this context, genetically modified transgenes compatible with opening up new opportunities unlimited many disorders. Recent advances in de novo protein design remodelling paving way new, more efficient targeted therapeutics. Using computational tools, expression cassette transgenic DNA can be split, miniaturized, shuffled or created from scratch to mediate transfer into cells. review, we highlight recent therapy focus on its use translational research. We summarize research therapy, an emphasis large (>4.8 kb) optimizing strategies applied by biomedical companies pipeline. critically discuss prospects treatment some emerging challenges. anticipate that continued novel tools will lead rapid basic studies.

Язык: Английский

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Red Light Optogenetics in Neuroscience

Frontiers in Cellular Neuroscience, Год журнала: 2022, Номер 15

Опубликована: Янв. 3, 2022

Optogenetics, a field concentrating on controlling cellular functions by means of light-activated proteins, has shown tremendous potential in neuroscience. It possesses superior spatiotemporal resolution compared to the surgical, electrical, and pharmacological methods traditionally used studying brain function. A multitude optogenetic tools for neuroscience have been created that, example, enable control action generation via ion channels. Other proteins brain, long-term potentiation or ablate specific subtypes neurons. In vivo applications, however, majority are operated with blue, green, yellow light, which all limited penetration biological tissues red light especially infrared light. This difference is significant, considering size rodent major research model Our review will focus utilization light-operated We first outline advantages studies. Then we provide brief overview systems new developments field. Finally, highlight different further facilitate use optogenetics

Язык: Английский

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Toward Optogenetic Hearing Restoration

Annual Review of Neuroscience, Год журнала: 2024, Номер 47(1), С. 103 - 121

Опубликована: Апрель 10, 2024

The cochlear implant (CI) is considered the most successful neuroprosthesis as it enables speech comprehension in majority of million otherwise deaf patients. In hearing by electrical stimulation auditory nerve, broad spread current from each electrode acts a bottleneck that limits transfer sound frequency information. Hence, there remains major unmet medical need for improving quality with CIs. Recently, optogenetic cochlea has been suggested an alternative approach restoration. Cochlear optogenetics promises to more information, hence hearing, light can conveniently be confined space activate nerve within smaller tonotopic ranges. this review, we discuss latest experimental and technological developments restoration outline remaining challenges en route clinical translation.

Язык: Английский

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Protein Design Accelerates the Development and Application of Optogenetic Tools

Computational and Structural Biotechnology Journal, Год журнала: 2025, Номер 27, С. 717 - 732

Опубликована: Янв. 1, 2025

Optogenetics has substantially enhanced our understanding of biological processes by enabling high-precision tracking and manipulation individual cells. It relies on photosensitive proteins to monitor control cellular activities, thereby paving the way for significant advancements in complex system research. Photosensitive play a vital role development optogenetics, facilitating establishment cutting-edge methods. Recent breakthroughs protein design have opened up opportunities develop protein-based tools that can precisely manipulate activities. These will significantly accelerate application optogenetic tools. This article emphasizes pivotal tools, offering insights into potential future directions. We begin providing an introduction historical fundamental principles followed exploration operational mechanisms key domains, which includes clarifying conformational changes they undergo response light, such as allosteric modulation dimerization processes. Building this foundation, we reveal enable creation even more sophisticated techniques.

Язык: Английский

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Is there an unmet medical need for improved hearing restoration?

EMBO Molecular Medicine, Год журнала: 2022, Номер 14(8)

Опубликована: Июль 14, 2022

Язык: Английский

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Utility of red‐light ultrafast optogenetic stimulation of the auditory pathway

EMBO Molecular Medicine, Год журнала: 2021, Номер 13(6)

Опубликована: Май 7, 2021

Optogenetic stimulation of spiral ganglion neurons (SGNs) in the ear provides a future alternative to electrical used current cochlear implants. Here, we employed fast and very variants red-light-activated channelrhodopsin (ChR) Chrimson (f-Chrimson vf-Chrimson) study their utility for optogenetic SGNs mice. The light requirements were higher vf-Chrimson than f-Chrimson, even when optimizing membrane expression by adding potassium channel trafficking sequences. time intensity coding single putative compared with acoustic clicks. enabled fire at near-physiological rates good temporal precision up 250 Hz stimulation. dynamic range SGN spike rate upon was narrower clicks but larger reported timing, on other hand, more comparable In conclusion, f-Chrimson are promising candidates auditory research

Язык: Английский

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Graded optogenetic activation of the auditory pathway for hearing restoration

Brain stimulation, Год журнала: 2023, Номер 16(2), С. 466 - 483

Опубликована: Янв. 23, 2023

Язык: Английский

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Fast Photoswitchable Molecular Prosthetics Control Neuronal Activity in the Cochlea

Journal of the American Chemical Society, Год журнала: 2022, Номер 144(21), С. 9229 - 9239

Опубликована: Май 18, 2022

Artificial control of neuronal activity enables the study neural circuits and restoration functions. Direct, rapid, sustained photocontrol intact neurons could overcome limitations established electrical stimulation such as poor selectivity. We have developed fast photoswitchable ligands glutamate receptors (GluRs) to enable in auditory system. The new induced photocurrents untransfected upon covalently tethering endogenous GluRs activating them reversibly with visible light pulses a few milliseconds. As proof concept these molecular prostheses, we applied ultrafast synapses cochlea that encode sound provide input brain. This drug-based method afforded optical adult gerbils at hundreds hertz without genetic manipulation would be required for their optogenetic control. indicates are also applicable spatiotemporal spiking interneurons

Язык: Английский

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Applications and challenges of rhodopsin-based optogenetics in biomedicine

Frontiers in Neuroscience, Год журнала: 2022, Номер 16

Опубликована: Сен. 23, 2022

Optogenetics is an emerging bioengineering technology that has been rapidly developed in recent years by cross-integrating optics, genetic engineering, electrophysiology, software control, and other disciplines. Since the first demonstration of millisecond neuromodulation ability channelrhodopsin-2 (ChR2), application optogenetic basic life science research progressed, especially neurobiology, which driven development discipline. As tool protein, microbial rhodopsins have continuously explored, modified, optimized, with many variants becoming available, structural characteristics functions are highly diversified. Their applicability broadened, encouraging more researchers clinicians to utilize optogenetics research. In this review, we summarize species variant types most important class proteins techniques, rhodopsins, review current applications based on rhodopsin qualitative light biology fields. We also challenges facing technology, ultimately provide in-depth technical reference support translational clinical

Язык: Английский

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Channelrhodopsin fluorescent tag replacement for clinical translation of optogenetic hearing restoration

Molecular Therapy — Methods & Clinical Development, Год журнала: 2023, Номер 29, С. 202 - 212

Опубликована: Март 21, 2023

Sensory restoration by optogenetic neurostimulation provides a promising future alternative to current electrical stimulation approaches. So far, channelrhodopsins (ChRs) typically contain C-terminal fluorescent protein (FP) tag for visualization that potentially poses an additional risk clinical translation. Previous work indicated reduction of efficacy upon FP removal. Here, we further optimized the fast-gating, red-light-activated ChR f-Chrimson achieve efficient in absence FP. Upon removal, observed massive amplitude photocurrents transfected cells vitro and optogenetically evoked activity adeno-associated virus (AAV) vector-transduced auditory nerve mice vivo. Increasing AAV vector dose restored but was confounded neural loss. Of various modifications, found replacement Kir2.1 trafficking sequence (TSKir2.1) best restore both with only mild loss few months after dosing. In conclusion, consider f-Chrimson-TSKir2.1 be candidate translation such as optical cochlear implants.

Язык: Английский

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