Skip to main content
Return to AAV Results

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

Addgene #213936-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.0b1tpc
Abstract
We injected interneuron enhancer, BiPVe4, 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.0b1tpc
AAV_BiPVe4_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 in S1 L2/3. Imaged on ZEISS Axioscan 7 Slidescanner at 20x magnification.
Caption: AAV_BiPVe4_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 in S1 L2/3. Imaged on ZEISS Axioscan 7 Slidescanner at 20x magnification.
AAV_BiPVe4_dTomato_nlsdTomato (AAV PHP.eB) replicate 2 was injected into the retro-orbital sinus of a 4 week old mouse. RNAscope stain in L2/3 of S1 with probes for dTomato (red) and Syt2 (green), showing clear fluorescent reporter expression indicating enhancer activity. Lack of colocalization with Syt2 (colocalized cells in yellow), which is a marker for PV basket, but not PV chandelier cells. Imaged on ZEISS Axio Imager Z2 Upright Confocal Microscope at 20x magnification.
Caption: AAV_BiPVe4_dTomato_nlsdTomato (AAV PHP.eB) replicate 2 was injected into the retro-orbital sinus of a 4 week old mouse. RNAscope stain in L2/3 of S1 with probes for dTomato (red) and Syt2 (green), showing clear fluorescent reporter expression indicating enhancer activity. Lack of colocalization with Syt2 (colocalized cells in yellow), which is a marker for PV basket, but not PV chandelier cells. Imaged on ZEISS Axio Imager Z2 Upright Confocal Microscope at 20x magnification.

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

Vector Used
pAAV_BiPVe4_dTomato_nlsdTomato
Addgene #213936-PHPeB

Virus & Injection

Virus

Virus Name
pAAV_BiPVe4_dTomato_nlsdTomato
Serotype
PHP.eB
Source
Addgene
Catalog Number
213936-PHPeB
Promoter
Other
Other Promoter
BiPVe4 (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 rep 1: Original titer of 6.55E+12 vg/mL. Diluted with PBS1X to achieve 2.00E+11 vg/animal in an 80 µL injection For rep 2: Original titer of 6.42E+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 BiPVe4 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
Mix of native fluorescence and RNAscope
Assay & Results
2 RO 4 weeks BiPVe4 replicates showing clear labeling of PV cells, especially in L2-3. Rep 1 shows one image with native flourescence and another with the native signal overlayed with DAPI. Rep 2 is from RNAscope, with probes used for dTomato (red/violet) and Syt2 (green). Results show BiPVe4-positive cells do not really colocalize with Syt2, a marker for basket cells.
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