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AAV pCAG-FLEX-tdTomato-WPRE in Mouse (AAV9)

Addgene #51503-AAV9

Data Submitted By
Kyle Lillis
Submission Date
July 10, 2019
Publication Date
July 10, 2019 (modified May 16, 2023)
Abstract
Not provided
This image was captured using a custom inverted single-photon widefield fluorescence microscope (shown is the red channel only). An Olympus 20X 0.95 NA objective was used to acquire a 5x5 grid of z-stacks. The image shown is a stitched maximum z-projection spanning essentially the entire volume of the slice.
Caption: This image was captured using a custom inverted single-photon widefield fluorescence microscope (shown is the red channel only). An Olympus 20X 0.95 NA objective was used to acquire a 5x5 grid of z-stacks. The image shown is a stitched maximum z-projection spanning essentially the entire volume of the slice.

Any further use of this image requires permission from the lab who submitted the data.

Vectors Used
AAV pCAG-FLEX-tdTomato-WPRE
Addgene #51503-AAV9
pGP-AAV-syn-jGCaMP7f-WPRE (AAV9)
Addgene #104488-AAV9

Virus & Injection

Virus 1

Virus Name
AAV pCAG-FLEX-tdTomato-WPRE
Serotype
AAV9
Source
Addgene
Catalog Number
51503-AAV9
Promoter
CAG
Cargo Type
Control
Injection Titer / Dose
1.00E+13 GC/mL
Injection Volume
5 μL
Injection Rate
Unknown, manual injection
Injector Material
Not provided
Injection Site / Route
Intracerebroventricular (ICV), NeuroNames ID : 2497
Additional Details
Imaging was performed on hippocampal organotypic slice cultures. Interneurons expressed the construct.
View in Addgene Catalog

Virus 2

Virus Name
pGP-AAV-syn-jGCaMP7f-WPRE (AAV9)
Serotype
AAV9
Source
Addgene
Catalog Number
104488-AAV9
Promoter
Synapsin
Injection Titer / Dose
1.00E+13 GC/mL
Injection Volume
5 uL
Injection Rate
Unknown (manual)
Injector Material
Not provided
Injection Site / Route
Cerebrospinal fluid (CSF): Intracerebroventricular (ICV)
Additional Details
pGP-AAV-syn-jGCaMP7f-WPRE (AAV9) was co-injected but is not shown in the image.
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
Dlx5/6-cre
Source
Jackson Labs
Stock Number
008199
Age at Injection
P1
Time After Injection
1 week
Detection Method
Direct fluorescence
Assay & Results
ICV injections at P0-P1 of AAV9 works very well at producing widespread expression throughout the brain, particularly in the hippocampi. The tool was intended to generate red fluorescence in all dlx-cre-expressing interneurons. The pattern of expression matched that expected and was very bright.
Tips & Comments
Vectors with the CAG promoter make for very bright expression - I would probably use a smaller volume of AAV in the future, for the sake of minimizing the impact of injection.