GEARBOCS: An Adeno Associated Virus Tool for In Vivo Gene Editing in Astrocytes DOI Open Access

Dhanesh Sivadasan Bindu,

Justin T Savage, Nicholas Brose

et al.

Published: Dec. 11, 2024

CRISPR/Cas9-based genome engineering enables rapid and precise gene manipulations in the CNS. Here, we developed a non-invasive astrocyte-specific method utilizing single AAV vector, which named GEARBOCS (Gene Editing AstRocytes Based On CRISPR/Cas9 System). We verified GEARBOCS’ specificity to mouse cortical astrocytes demonstrated its utility for three types of manipulations: knockout (KO); tagging (TagIn); reporter knock-in (GeneTrap) strategies. Next, deployed two test cases. First, determined that are necessary source synaptogenic factor Sparcl1 thalamocortical synapse maintenance primary visual cortex. Second, express synaptic vesicle associated Vamp2 protein found it is required maintaining excitatory inhibitory numbers These results show strategy provides fast efficient means study astrocyte biology vivo .

Language: Английский

GEARBOCS: An Adeno Associated Virus Tool forIn VivoGene Editing in Astrocytes DOI Creative Commons

Dhanesh Sivadasan Bindu,

Justin T Savage, Nicholas Brose

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Jan. 19, 2023

Summary CRISPR/Cas9-based genome engineering enables rapid and precise gene manipulations in the CNS. Here, we developed a non-invasive astrocyte-specific method utilizing single AAV vector, which named GEARBOCS (Gene Editing AstRocytes Based On CRISPR/Cas9 System). We verified GEARBOCS’ specificity to mouse cortical astrocytes demonstrated its utility for three types of manipulations: knockout (KO); tagging (TagIn); reporter knock-in (GeneTrap) strategies. Next, deployed two test cases. First, determined that are necessary source synaptogenic factor Sparcl1 thalamocortical synapse maintenance primary visual cortex. Second, express synaptic vesicle associated Vamp2 protein found it is required maintaining excitatory inhibitory numbers These results show strategy provides fast efficient means study astrocyte biology vivo . Motivation Astrocytes indispensable brain development, function, health. However, molecular tools function have been largely limited genetically modified mice. editing within vector astrocytes. designed optimized this easy-to-use viral tool understand expression, localization

Language: Английский

Citations

4
Astrocytic modulation of population encoding in mouse visual cortex via GABA transporter 3 revealed by multiplexed CRISPR/Cas9 gene editing DOI Creative Commons
Ji Ho Park, Grayson O. Sipe, Xin Tang

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 7, 2024

Summary Astrocytes, which are increasingly recognized as pivotal constituents of brain circuits governing a wide range functions, express GABA transporter 3 (Gat3), an astrocyte-specific responsible for maintenance extra-synaptic levels. Here, we examined the functional role Gat3 in astrocyte-mediated modulation neuronal activity and information encoding. First, developed multiplexed CRISPR construct applicable effective genetic ablation visual cortex adult mice. Using vivo two-photon calcium imaging neurons knockout mice, observed changes spontaneous visually driven single response properties such magnitudes trial-to-trial variability. exerted pronounced influence on population-level activity, altering dynamics populations impairing their ability to accurately represent stimulus information. These findings demonstrate that astrocytes profoundly shapes sensory encoding capacity networks within cortex.

Language: Английский

Citations

0
GEARBOCS: An Adeno Associated Virus Tool for In Vivo Gene Editing in Astrocytes DOI Open Access

Dhanesh Sivadasan Bindu,

Justin T Savage, Nicholas Brose

et al.

Published: Dec. 11, 2024

CRISPR/Cas9-based genome engineering enables rapid and precise gene manipulations in the CNS. Here, we developed a non-invasive astrocyte-specific method utilizing single AAV vector, which named GEARBOCS (Gene Editing AstRocytes Based On CRISPR/Cas9 System). We verified GEARBOCS’ specificity to mouse cortical astrocytes demonstrated its utility for three types of manipulations: knockout (KO); tagging (TagIn); reporter knock-in (GeneTrap) strategies. Next, deployed two test cases. First, determined that are necessary source synaptogenic factor Sparcl1 thalamocortical synapse maintenance primary visual cortex. Second, express synaptic vesicle associated Vamp2 protein found it is required maintaining excitatory inhibitory numbers These results show strategy provides fast efficient means study astrocyte biology vivo .

Language: Английский

Citations

0
GEARBOCS: An Adeno Associated Virus Tool for In Vivo Gene Editing in Astrocytes DOI Open Access

Dhanesh Sivadasan Bindu,

Justin T Savage, Nicholas Brose

et al.

Published: Dec. 11, 2024

CRISPR/Cas9-based genome engineering enables rapid and precise gene manipulations in the CNS. Here, we developed a non-invasive astrocyte-specific method utilizing single AAV vector, which named GEARBOCS (Gene Editing AstRocytes Based On CRISPR/Cas9 System). We verified GEARBOCS’ specificity to mouse cortical astrocytes demonstrated its utility for three types of manipulations: knockout (KO); tagging (TagIn); reporter knock-in (GeneTrap) strategies. Next, deployed two test cases. First, determined that are necessary source synaptogenic factor Sparcl1 thalamocortical synapse maintenance primary visual cortex. Second, express synaptic vesicle associated Vamp2 protein found it is required maintaining excitatory inhibitory numbers These results show strategy provides fast efficient means study astrocyte biology vivo .

Language: Английский

Citations

0