Evidence map›Paper›PMID 40700012›Full record

ArticleSTAR protocols2025

Protocol for CRISPR-based manipulation and visualization of endogenous α-synuclein in cultured mouse hippocampal neurons.

Leonardo A Parra-Rivas, Rohan Sharma, Trinity E Rust, Hannah O Bazick, Jared Carlson-Stevermer, Mark J Zylka, Yuki Ogawa, Subhojit Roy

Abstract read
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In one paragraph

Article in STAR protocols, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors.

Leonardo A Parra-RivasDepartment of Pathology, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, USA; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA.
Rohan SharmaDepartment of Pathology, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, USA.
Trinity E RustDepartment of Cell Biology and Physiology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Hannah O BazickUNC Neuroscience Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Jared Carlson-StevermerSerotiny Inc., 329 Oyster Point Boulevard, 3rd Floor, South San Francisco, CA 94080, USA.
Mark J ZylkaDepartment of Cell Biology and Physiology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA; UNC Neuroscience Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA; Carolina Institute for Developmental Disabilities, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Yuki OgawaDepartment of Neuroscience, Baylor College of Medicine, Houston, TX, USA.
Subhojit RoyDepartment of Pathology, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, USA; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA; Department of Neuroscience, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, USA. Electronic address: sroy@ucsd.edu.

Funding

UCSD Shiley-Marcos Alzheimer's Disease Research Center P30P30AG062429 · NIA · UNIVERSITY OF CALIFORNIA, SAN DIEGO · PI DOUGLAS R GALASKO · 2019 to 2026
$34.9M
Pathophysiologic roles of alpha-synuclein at the synapseR01NS111978 · NINDS · UNIVERSITY OF CALIFORNIA, SAN DIEGO · PI ROY, SUBHOJIT · 2019 to 2023
$2.4M
NIA NIH HHS P30 AG062429NINDS NIH HHS R01 NS111978
6 · The paper itself

Abstract

CRISPR-Cas9 technology enables acute gene knockdown and endogenous tagging to study single-synapse function. Here, we present a protocol for depleting alpha-synuclein (α-syn) or visualizing native α-syn with an endogenously inserted fluorescent tag in cultured mouse hippocampal neurons. We describe detailed steps, including CRISPR design, virus packaging/transduction (delivery), and validation of on-/off-target editing. This protocol should be useful for assigning precise function to contentious synaptic proteins and for visualizing protein trafficking without overexpression in cultured hippocampal neurons-an established model system for synaptic biology. For complete details on the use and execution of this protocol, please refer to Parra-Rivas et al.

Indexed as

alpha-SynucleinCRISPR-Cas SystemsHippocampusNeuronsAnimalsCells, CulturedGene EditingMicealpha-SynucleinCell BiologyCell cultureCRISPRMicroscopyMolecular BiologyNeuroscience

Identifiers

PMID40700012
PMCPMC12305205

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.