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Related Experiment Video

Updated: Sep 13, 2025

Widespread Transduction of Mouse Neocortical Neurons by Subarachnoid Injection of AAV2
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AAV vectors for specific and efficient gene expression in microglia.

Ryo Aoki1, Ayumu Konno2, Nobutake Hosoi1

  • 1Department of Neurophysiology & Neural Repair, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan.

Cell Reports Methods
|July 31, 2025
PubMed
Summary

Researchers developed a novel adeno-associated virus (AAV) vector for precise gene delivery to microglia. This tool enhances microglial research and potential therapies for neurological diseases.

Keywords:
AAVCP: neuroscienceCa(2+) imagingGCaMPIba1RNA interferenceWPREadeno-associated viruscerebral cortexmicroRNAmicroglia

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Gene Therapy

Background:

  • Microglia are key players in neurological diseases like Alzheimer's and stroke.
  • Efficient and specific gene delivery to microglia remains a significant challenge.
  • Targeting microglia is crucial for developing new therapeutic strategies.

Purpose of the Study:

  • To engineer an adeno-associated virus (AAV) vector for highly specific and efficient microglial gene delivery.
  • To validate the vector's performance in terms of specificity, efficiency, and long-term expression.
  • To demonstrate the vector's utility for monitoring microglial activity in real-time.

Main Methods:

  • Development of a custom AAV vector utilizing the mIba1 promoter, GFP reporter, miRNA target sequences (miR.Ts), WPRE, and poly(A) signal.
  • Strategic placement of miR.Ts to minimize off-target expression.
  • Administration of the AAV vector, including a blood-brain barrier-penetrant AAV-9P31 capsid variant, via intravenous injection.

Main Results:

  • Achieved over 90% specificity and >60% efficiency for microglia-specific gene expression 3 weeks post-administration.
  • Demonstrated suppression of non-microglial expression by positioning miR.Ts flanking the WPRE.
  • Enabled GCaMP expression for real-time monitoring of microglial calcium dynamics.
  • Intravenous administration of the AAV-9P31 variant resulted in widespread, selective microglial expression across the brain.

Conclusions:

  • The developed AAV vector provides a versatile tool for achieving long-term, highly specific, and efficient gene expression in microglia.
  • This technology significantly advances the potential for microglial research and therapeutic applications in neurological disorders.
  • The vector enables precise genetic manipulation and functional studies of microglia in vivo.