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Cited by 58Virus-based gene therapy researchNeuroinflammation and Neurodegeneration MechanismsRNA Interference and Gene Delivery

Development of microglia‐targeting adeno‐associated viral vectors as tools to study microglial behavior in vivo

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Okada, Yukihiro, Hosoi, Nobutake, Matsuzaki, Yasunori, Fukai, Yuuki, Hiraga, Akito, Nakai, Junichi, Nitta, Keisuke, Shinohara, Yoichiro +2 more·2022·Communications Biology
DOI 10.1038/s42003-022-04200-3PMC PMC9652230
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Abstract

Here we describe the microglia-targeting adeno-associated viral (AAV) vectors containing a 1.7-kb putative promoter region of microglia/macrophage-specific ionized calcium-binding adaptor molecule 1 ( Iba1 ), along with repeated miRNA target sites for microRNA (miR)-9 and miR-129-2-3p. The 1.7-kb genomic sequence upstream of the start codon in exon 1 of the Iba1 ( Aif1 ) gene, functions as microglia preferential promoter in the striatum and cerebellum. Furthermore, ectopic transgene expression in non-microglial cells is markedly suppressed upon adding two sets of 4-repeated miRNA target sites for miR-9 and miR-129-2-3p, which are expressed exclusively in non-microglial cells and sponged AAV-derived mRNAs. Our vectors transduced ramified microglia in healthy tissues and reactive microglia in lipopolysaccharide-treated mice and a mouse model of neurodegenerative disease. Moreover, live fluorescent imaging allowed the monitoring of microglial motility and intracellular Ca 2+ mobilization. Thus, microglia-targeting AAV vectors are valuable for studying microglial pathophysiology and therapies, particularly in the striatum and cerebellum.

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  • The IBA1 promoter has been validated extensively for microglial specificity, with reported transduction specificity of up to 95% in resident microglia.

    “The IBA1 promoter used in this study has been validated extensively for microglial specificity, with reported transduction specificity of up to 95% in resident microglia. 33 , 34 , 35”

    From Aberrant phase separation from a rare <i>ABI3</i> mutation drives microglial dysfunction and Alzheimer's risk (2026) · cites this paper as [34]

    The Results section on Iba1-promoter targeting (page unavailable) reports preferential microglial expression in the striatum (~69%) and cerebellum (~86%), but almost no microglial specificity in the cerebral cortex (~2%). A later Results section reports that a vector combining the promoter with miRNA target sequences transduced almost exclusively microglia in the striatum and cerebellum; it does not attribute 95% specificity to the promoter alone. The Discussion (page unavailable) also cautions that the promoter acts in other myeloid cells. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • The IBA1 promoter paired with an artificial miRNA scaffold is a well-established strategy for microglia-specific gene manipulation.

    “This promoter, paired with an artificial miRNA scaffold embedded in the miR‐RNA backbone, is a well‐established strategy for microglia‐specific gene manipulation. 33 , 34 , 35”

    From Aberrant phase separation from a rare <i>ABI3</i> mutation drives microglial dysfunction and Alzheimer's risk (2026) · cites this paper as [34]

    The Methods (Vector construction; page unavailable) describe pairing the Iba1 promoter with miRNA target sequences, and the Results report microglial targeting with the resulting AAV vector. They do not describe an artificial miRNA scaffold embedded in a miR-RNA backbone. The Discussion also cautions that the Iba1 promoter is not strictly microglia-specific, so the paper does not establish the claimed pairing as a well-established strategy. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Cited by

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Authors
Okada, Y., Hosoi, N., Matsuzaki, Y., Fukai, Y., Hiraga, A., Nakai, J., Nitta, K., Shinohara, Y., Konno, A., & Hirai, H.
Affiliations
  • Gunma University (JP)
  • Tohoku University (JP)
Corresponding author
Hirai, Hirokazu
ORCID
Venue
Communications Biology
Publisher
Springer Science and Business Media LLC
Published
2022-11-11
Volume
5
Issue
1
Pages
1224-1224
ISSN
2399-3642 (ISSN-L)
DOI
10.1038/s42003-022-04200-3
PMC
PMC9652230
OpenAlex
W4309046731
Cited by
58
References
8
Citations by year
2026: 13 · 2025: 21 · 2024: 19 · 2023: 5
FWCI
6.79
Type
journal article
Language
en
Open access
Licence
Locations
  • Communications Biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Japan Agency for Medical Research and Development (19K06899, JP21dm0207111, JP19dm0207057, 20dm0207057h0004, 22K06454)
  • Japan Society for the Promotion of Science (15K18330, 16K15477, 22K06454, 19K06899, 20K06906, 18H02521, JP21dm0207111, JP20dm0207057, 15H04254)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
microRNA binding sites, miR-9, microglial motility, calcium mobilization, reactive microglia, lipopolysaccharide, neurodegenerative diseases, striatum, cerebellum, transgene expression
MeSH terms
Animals, Lipopolysaccharides, Microglia, Transgenes, Neurodegenerative Diseases, MicroRNAs, Mice

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