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

Addgene #218792-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.9p2jpw
Abstract
We injected interneuron enhancer, BiLAMP5e, 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.9p2jpw
pAAV_BiLAMP5e3_dTomato_nlsdTomato (AAV PHP.eB) replicate 1 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal snap of the cortex showing clear native fluorescent reporter expression (red) indicating enhancer activity across S1 layers. Imaged on ZEISS Axio Imager M2 upright microscope at 5x magnification.
Caption: pAAV_BiLAMP5e3_dTomato_nlsdTomato (AAV PHP.eB) replicate 1 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal snap of the cortex showing clear native fluorescent reporter expression (red) indicating enhancer activity across S1 layers. Imaged on ZEISS Axio Imager M2 upright microscope at 5x magnification.
pAAV_BiLAMP5e3_dTomato_nlsdTomato (AAV PHP.eB) replicate 2 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal snap of the cortex showing clear native fluorescent reporter expression (red) indicating enhancer activity across S1 layers. Imaged on ZEISS Axio Imager M2 upright microscope at 5x magnification.
Caption: pAAV_BiLAMP5e3_dTomato_nlsdTomato (AAV PHP.eB) replicate 2 was injected into the retro-orbital sinus of a 4 week old mouse. Coronal snap of the cortex showing clear native fluorescent reporter expression (red) indicating enhancer activity across S1 layers. Imaged on ZEISS Axio Imager M2 upright microscope at 5x magnification.

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

Vector Used
pAAV_BiLAMP5e3_dTomato_nlsdTomato
Addgene #218792-PHPeB

Virus & Injection

Virus

Virus Name
pAAV_BiLAMP5e3_dTomato_nlsdTomato
Serotype
PHP.eB
Source
Addgene
Catalog Number
218792-PHPeB
Promoter
Other
Other Promoter
BiLAMP5e3 (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 5.84E+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 BiLAMP5e3 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 BiLAMP5 replicates showing clear viral labeling of Lamp5 cells across L2-6. Images shows native dTomato reporter fluorescence (red/violet).
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