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CRISPR/Cas9 constructs can be delivered by in utero electroporation to knock out a gene of interest in neurons of the developing brain in wild-type rodents. This approach allows for high-throughput genetic screening, circuit-specific gene knockout, and knockout cell phenotyping using sparse labeling within a wild-type in vivo context. Here we outline the methods and steps of designing guide RNAs in silico, cloning guide RNAs into plasmid backbones, and introducing these plasmids into the developing mouse cortex and hippocampus.

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Gene Knockout in the Developing Brain of Wild-Type Rodents by CRISPR In Utero Electroporation

  • 实验方案

  • 发表日期：

    2025年

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  • Andrea J. Romanowski,
  • Ryan R. Richardson,
  • Celine Plachez,
  • Reha S. Erzurumlu,
  • Alexandros Poulopoulos

摘要

CRISPR/Cas9 constructs can be delivered by in utero electroporation to knock out a gene of interest in neurons of the developing brain in wild-type rodents. This approach allows for high-throughput genetic screening, circuit-specific gene knockout, and knockout cell phenotyping using sparse labeling within a wild-type in vivo context. Here we outline the methods and steps of designing guide RNAs in silico, cloning guide RNAs into plasmid backbones, and introducing these plasmids into the developing mouse cortex and hippocampus.