Widespread labeling and genomic editing of the fetal central nervous system by in utero CRISPR AAV9-PHP.eB administration

Development. 2021 Jan 20;148(2):dev195586. doi: 10.1242/dev.195586.

Abstract

Efficient genetic manipulation in the developing central nervous system is crucial for investigating mechanisms of neurodevelopmental disorders and the development of promising therapeutics. Common approaches including transgenic mice and in utero electroporation, although powerful in many aspects, have their own limitations. In this study, we delivered vectors based on the AAV9.PHP.eB pseudo-type to the fetal mouse brain, and achieved widespread and extensive transduction of neural cells. When AAV9.PHP.eB-coding gRNA targeting PogZ or Depdc5 was delivered to Cas9 transgenic mice, widespread gene knockout was also achieved at the whole brain level. Our studies provide a useful platform for studying brain development and devising genetic intervention for severe developmental diseases.

Keywords: AAV9-PHP.eB; CRISPR; DEPDC5; Mouse; POGZ.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Central Nervous System / embryology*
  • Central Nervous System / metabolism*
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Dependovirus / metabolism*
  • Disease Susceptibility
  • Female
  • Fetus / metabolism*
  • GTPase-Activating Proteins / genetics
  • GTPase-Activating Proteins / metabolism
  • Gene Editing*
  • Gene Expression Regulation, Developmental
  • Gene Knockout Techniques
  • Gene Transfer Techniques
  • Male
  • Mice
  • Seizures / pathology
  • Staining and Labeling*

Substances

  • Depdc5 protein, mouse
  • GTPase-Activating Proteins