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CN1851 Characterization

Addgene #164450

Data Submitted By
Boaz Levi , Bosiljka Tasic and Jonathan Ting
Submission Date
June 16, 2023
Publication Date
June 27, 2023 (modified June 27, 2023)
DOI
https://doi.org/10.57733/addgene.l0iuyl
Funding
NIA/NIMH RF1MH114126, UF1MH128339
Abstract
Forebrain GABAergic neurons are labeled in mouse brain by transduction with CN1851 AAV of the PHP.eB serotype. It was injected at a concentration of 1.0E+11 viral genomes per animal into one month-old Ai14 Cre recombinase reporter mice via the retro-orbital route. The animal was perfused and the brain fixed in 4% paraformaldehyde four weeks post injection. The fixed brain tissue was then sectioned, slide mounted, and imaged for native tdTomato fluorescence. CN1851 has been characterized and was demonstrated to label forebrain GABAergic neurons in Graybuck et. al. (Neuron) 2021. https://doi.org/10.1016/j.neuron.2021.03.011.
Citation
CN1851 Characterization. Levi B, Tasic B, Ting J 2023. Addgene Report, https://doi.org/10.57733/addgene.l0iuyl
Caption: A sagittal section of a whole mouse brain is shown on the left, and cortex region is shown on the right (including VISp and PFLp, marked by yellow box). Forebrain GABAergic neurons are labeled in mouse brain by transduction with AAV-PHP.eB/CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA. The section was imaged for native tdTomato fluorescence on a Leica Aperio Versa slide scanner. CN1851 has been characterized and was demonstrated to label forebrain GABAergic neurons in Graybuck et. al. (Neuron) 2021. https://doi.org/10.1016/j.neuron.2021.03.011.
Image attribution: VGT Pipeline, Allen Institute for Brain Science

This report is made available under the Creative Commons Attribution 4.0 International License.

Vector Used
CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA
Addgene #164450

Virus & Injection

Virus

Virus Name
CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA
Serotype
PHP.eB
Source
Addgene
Catalog Number
164450
Virus Lot Number
v165478G
Promoter
Other
Other Promoter
hI56i
Cargo Type
Recombinase
Injection Titer / Dose
1.0 x 10e11 vg/animal
Injection Volume
90 ┬Ál
Injection Rate
N/A
Injector Material
31g insulin syringe
Tip Diameter
31
Coatings
N/A
Injection Time
N/A
Injection Site / Route
Retro-orbital injection
Additional Details
AAV-PHP.eB/CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA was injected at a concentration of 1.0 x 10e11 viral genomes per animal into one month-old Ai14 Cre recombinase reporter mice via the retro-orbital route.
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
Ai14(RCL-tdT): B6.CgGt(ROSA)26Sortm14(CAG-tdTomato)Hze/J
Source
Jackson Laboratory
Stock Number
007914
Age at Injection
1 month
Sex
Male
Time After Injection
4 Weeks
Detection Method
Native fluorescence
Assay & Results
The animal was perfused and the brain fixed with 4% paraformaldehyde four weeks post injection. The fixed brain tissue was then sectioned, slide mounted, and imaged for native tdTomato fluorescence on a Leica Aperio Versa slide scanner.
Target Cell Transduction
N/A
Functional Testing
iCre was functional as expected.