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Characterization of an enhancer-AAV aimed at targeting distinct interneuron subtypes

Addgene #213940-PHPeB

Data Submitted By
Elisabetta Furlanis , Min Dai , Brenda Leyva Garcia , Sara Wills , Jordane Dimidschstein , Gordon Fishell and Yating Wang
Lab Name
Gordon Fishell, Harvard Medical School
Submission Date
January 06, 2026
Publication Date
January 26, 2026 (modified January 26, 2026)
DOI
https://doi.org/10.57733/addgene.fhdkro
Abstract
We injected interneuron enhancer, BiPVe3, tagged with a dTomato reporter through retro-orbital injection routes into mice. After waiting 3 weeks to allow for infection, we processed the brains and looked at endogenous dTomato reporter expression to assess enhancer activity in the primary somatosensory cortex. This project took place as part of the Brain Initiative and we have uploaded our results here.
Citation
Characterization of an enhancer-AAV aimed at targeting distinct interneuron subtypes. Furlanis E, Dai M, Leyva Garcia B, Wills S, Dimidschstein J, Fishell G, Wang Y 2026. Addgene Report, https://doi.org/10.57733/addgene.fhdkro
pAAV_BiPVe3_dTomato_nlsdTomato (AAV PHP.eB) replicate 1 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal section showing clear native fluorescent reporter expression (violet) indicating enhancer activity, particularly across S1 layers 2–6. Imaged on ZEISS Axioscan 7 Slidescanner at 20x magnification.
Caption: pAAV_BiPVe3_dTomato_nlsdTomato (AAV PHP.eB) replicate 1 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal section showing clear native fluorescent reporter expression (violet) indicating enhancer activity, particularly across S1 layers 2–6. Imaged on ZEISS Axioscan 7 Slidescanner at 20x magnification.
pAAV_BiPVe3_dTomato_nlsdTomato (AAV PHP.eB) replicate 2 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal section showing clear native fluorescent reporter expression (red) indicating enhancer activity, particularly across S1 layers 2–6. Imaged on ZEISS Axioscan 7 Slidescanner at 20x magnification.
Caption: pAAV_BiPVe3_dTomato_nlsdTomato (AAV PHP.eB) replicate 2 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal section showing clear native fluorescent reporter expression (red) indicating enhancer activity, particularly across S1 layers 2–6. Imaged on ZEISS Axioscan 7 Slidescanner at 20x magnification.

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

Vector Used
pAAV_BiPVe3_dTomato_nlsdTomato
Addgene #213940-PHPeB

Virus & Injection

Virus

Virus Name
pAAV_BiPVe3_dTomato_nlsdTomato
Serotype
PHP.eB
Source
Addgene
Catalog Number
213940-PHPeB
Promoter
Other
Other Promoter
BiPVe3 (enhancer)
Cargo Type
Control
Injection Titer / Dose
2.00E+11 vg/animal
Injection Volume
80 µL
Injection Rate
N/A
Injector Material
BD Insulin Syringe 0.3 mL
Injection Time
N/A
Injection Site / Route
Retro-Orbital (RO) injection into the retro-orbital sinus of 4 week old mouse for systemic expression across the brain.
Additional Details
For both reps, original titer of 5.54E+12 vg/mL. Diluted with PBS1X to achieve 2.00E+11 vg/animal in an 80 µL injection
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
For both mice injected with BiPVe3 enhancer: C57BL/6J
Source
JAX
Stock Number
JAX:000664
Age at Injection
4 weeks
Sex
Mixed
Time After Injection
3 weeks before perfusion
Detection Method
Native fluorescence
Assay & Results
2 RO 4 weeks BIPVe3 replicates showing clear labeling of PV cells across L2-6. Both replicates have an image showing native dTomato reporter fluorescence (red/violet), and another with the native signal overlayed with DAPI.
Tips & Comments
Please see the original publication for more details: Furlanis, E., Dai, M., Leyva Garcia, B., Tran, T., Vergara, J., Pereira, A., Gorissen, B. L., Wills, S., Vlachos, A., Hairston, A., Dwivedi, D., Du, S., McMahon, J., Huang, S., Morabito, A., Vazquez, A., Kim, S., Lee, A. T., Chang, E. F., Razzaq, T., … Wang, Y. (2025). An enhancer-AAV toolbox to target and manipulate distinct interneuron subtypes. Neuron, 113(10), 1525–1547.e15. https://doi.org/10.1016/j.neuron.2025.05.002