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CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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HIV-1 mRNA knockdown with CRISPR/CAS9 enhances neurocognitive function.

Kristen A McLaurin1,2, Hailong Li1, Kamel Khalili3

  • 1Cognitive and Neural Science Program, Department of Psychology, Barnwell College, University of South Carolina, 1512 Pendleton Street, Columbia, SC, 29208, USA.

Journal of Neurovirology
|February 15, 2024
PubMed
Summary

CRISPR/Cas9 gene editing effectively reduced HIV-1 mRNA in mixed glia, showing promise for treating HIV-associated neurocognitive disorders (HAND). This approach restored temporal processing, suggesting a potential therapeutic strategy even without full viral eradication.

Keywords:
Cell cultureGene editingIn situ hybridizationMicrogliaPrepulse inhibition

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Area of Science:

  • Neuroscience
  • Genetics
  • Virology

Background:

  • HIV-1 establishes persistent reservoirs in the central nervous system (CNS) within mixed glia.
  • Gene editing, particularly CRISPR/Cas9, offers potential for HIV-1 reservoir elimination.
  • The efficacy of CRISPR/Cas9 in removing HIV-1 proteins from mixed glia remains unevaluated.

Purpose of the Study:

  • To evaluate the efficacy of adeno-associated virus 9 (AAV9)-CRISPR/Cas9 gene editing in eliminating HIV-1 messenger RNA (mRNA) from cortical mixed glia.
  • To assess the impact of CRISPR/Cas9 treatment on neurobehavioral outcomes related to HIV-1-associated neurocognitive disorders (HAND).

Main Methods:

  • In vitro: Mixed glia from HIV-1 transgenic (Tg) rats were treated with varying doses of AAV9-CRISPR/Cas9.
  • In vivo: HIV-1 Tg rats received retro-orbital inoculation of AAV9-CRISPR/Cas9.
  • HIV-1 mRNA levels were quantified using in situ hybridization.
  • Temporal processing was assessed to evaluate neurocognitive function.

Main Results:

  • In vitro, a dose-dependent decrease in HIV-1 mRNA was observed in a subset of treated mixed glia.
  • In vivo, CRISPR/Cas9 treatment resulted in significant excision (approx. 53.2%) of HIV-1 mRNA in the medial prefrontal cortex.
  • CRISPR/Cas9 treatment partially restored the developmental trajectory of temporal processing.

Conclusions:

  • Mixed glia are susceptible to gene editing interventions.
  • AAV9-CRISPR/Cas9 demonstrates potential as a therapeutic strategy for HAND by reducing HIV-1 mRNA.
  • CRISPR/Cas9 may offer clinical benefits for HAND even without complete viral genome eradication.