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Updated: Apr 25, 2026

Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

Published on: January 3, 2015

Efficient, complete deletion of synaptic proteins using CRISPR.

Salvatore Incontro¹, Cedric S Asensio², Robert H Edwards²

¹Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA.

|August 27, 2014

Summary

This summary is machine-generated.

CRISPR/Cas9 gene editing effectively inactivated NMDA and AMPA receptor function in postmitotic neurons. This powerful new method allows researchers to study synaptic protein functions by precisely deleting target genes.

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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Area of Science:

Neuroscience
Molecular Biology
Genetics

Background:

Understanding protein function often involves studying gene deletion effects.
Traditional methods like homologous recombination and RNA interference (RNAi) have limitations.
CRISPR/Cas9 gene editing offers a novel approach for gene inactivation.

Purpose of the Study:

To evaluate the efficacy of CRISPR/Cas9 in inactivating synaptic protein function in postmitotic neurons.
To target the GluN1 subunit of NMDA receptors and the GluA2 subunit of AMPA receptors.

Main Methods:

Utilized CRISPR/Cas9 gene editing technology.
Targeted two key synaptic proteins: NMDA receptor (GluN1 subunit) and AMPA receptor (GluA2 subunit).
Applied the system in hippocampal slice cultures derived from postmitotic neurons.

Main Results:

CRISPR/Cas9 expression led to complete elimination of NMDA receptor function.
CRISPR/Cas9 expression resulted in complete elimination of GluA2 subunit function.
Demonstrated successful gene inactivation in postmitotic neurons.

Conclusions:

CRISPR/Cas9 is a potent tool for functional studies of synaptic proteins.
This method enables precise gene inactivation in challenging neuronal cell types.
Facilitates research into the roles of specific synaptic proteins in neuronal function.