The Most Comprehensive Collection of Gene Delivery Tools for Brain Endothelial Cells

2025: rAAV-miniBEND – Promoter Optimization + Ultra-Large Payload, Focusing on Precise Disease Modeling and Editing

The AAV-miniBEND system combines the smallest promoter isolated from the mouse Tek gene and optimized cis-regulatory elements to achieve specific gene expression in mouse and rat brain endothelial cells.

Features:

Figure 7: Optimization of rAAV-miniBEND promoter and high-targeting of endothelial cells
 

Application Cases

1.Cerebral Cavernous Malformation (CCM)

Mouse: Adult mouse
Virus: rAAV-PHP.eB-miniBEND-MAP3K3I441M
Injection Protocol: Stereotaxic injection into the cortex, ~1.5-5×10⁹ vg/mouse; intravenous injection ~1.5-5×10¹¹ vg/mouse, expression for 2-3 weeks
Results: Using the rAAV-miniBEND expression system, human MAP3K3 gene with I441M point mutation was delivered to the mouse brain endothelial cells. Intravenous injection induced widespread brain lesions, while local injection established focal lesions, successfully generating a sporadic CCM mouse model. The model displayed pathological features consistent with clinical grade IV CCM, including popcorn-like hemorrhage, blood-brain barrier leakage, and vascular cavernous dilation. MRI and various histological stains confirmed these features. Compared to traditional AAV serotypes, the rAAV-miniBEND system demonstrated superior targeting efficiency, making it ideal for in vivo delayed imaging studies of CCM lesions.

Figure 8: Establishing a CCM model using the rAAV-miniBEND system

 

2.Brain Arteriovenous Malformation (AVM) Modeling

Mouse: 30-50 day-old Braf-CAfl/fl mouse
Virus: rAAV-PHP.eB-miniBEND-Cre
Injection Protocol: Stereotaxic injection into the cortex or hippocampus, ~1.5-5×10⁹ vg/mouse; expression for 2 weeks before detection
Results: Local delivery of Cre recombinase into Braf-CAfl/fl mice specifically activated the BrafV600E oncogenic mutation, successfully constructing a rapidly progressing brain arteriovenous malformation (bAVM) mouse model in a few weeks. The model reproduced the core pathological features of human bAVM, including brain hemorrhage, edema, abnormally dilated malformed blood vessels, tissue necrosis, and blood-brain barrier disruption. The model mice exhibited clinical symptoms such as hemiplegia, seizures, and a high mortality rate. Further studies revealed vascular dilation, hemodynamic changes, endothelial-mesenchymal transition, and key arteriovenous shunting structures, providing powerful tools for understanding bAVM pathogenesis and drug screening.

Figure 9: Modeling bAVM using the rAAV-miniBEND system

Summary

From 2016 to 2025, AAV targeting technology for brain endothelial cells has achieved four key breakthroughs:
The targeting range has expanded from focusing on capillaries to covering the entire vascular segment, with cross-species compatibility.
Specificity has improved from brain-specific targeting to minimal off-target effects in peripheral organs.
Functionality has advanced from simple transduction to gene editing, repeated dosing, and disease modeling.

These tools not only address the technical bottlenecks in neurovascular research but also drive the development of gene therapies for diseases such as cerebrovascular malformations and stroke sequelae.

Summary of Viral Strategies for Targeting Brain Endothelial Cells:

References
[1]Körbelin J, Dogbevia G, Michelfelder S, et al. A brain microvasculature endothelial cell-specific viral vector with the potential to treat neurovascular and neurological diseases. EMBO Mol Med. 2016;8(6):609-625.
[2]Krolak T, Chan KY, Kaplan L, et al. A High-Efficiency AAV for Endothelial Cell Transduction Throughout the Central Nervous System. Nat Cardiovasc Res. 2022;1(4):389-400.  
[3]Chen X, Wolfe DA, Bindu DS, et al. Functional gene delivery to and across brain vasculature of systemic AAVs with endothelial-specific tropism in rodents and broad tropism in primates. Nat Commun. 2023;14(1):3345.
[4]Li JL, Bi Z, Chen XJ, et al. A targeted vector for brain endothelial cell gene delivery and cerebrovascular malformation modelling. Nat Biomed Eng. Published online October 29, 2025.