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Neuroscience JoVE (Open Access) Citable · DOI

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

DOI: 10.3791/3024-v
What you'll learn
  • ✓ Prepare capillaries and micro-electrodes for in utero electroporation
  • ✓ Execute surgical injection and electroporation in developing mouse brain
  • ✓ Achieve spatially and temporally controlled plasmid DNA transfection in vivo
Protocol

A gene transfer method into the developing mouse brain is described by using a unique surgical method and special shape of electrodes. This unique technique allows transfection of plasmid DNA temporally and spatially, which will aid many neuroscientists in studying brain development.

Difficulty
advanced
Total time
~45–60 min per mouse (surgical procedure plus recovery)
Model organism
Mouse (embryonic in utero)
Biosafety
BSL-1

Steps

1
Prepare capillaries and micro-electrodes

Fabricate specialized capillaries and electrode shapes required for precise plasmid DNA delivery. This preparation ensures proper spatial targeting and electrical contact with developing neural tissue.

▶ 02:08
2
Prepare plasmid DNA for transfection

Isolate and prepare plasmid DNA constructs at appropriate concentration for electroporation. Quality and purity of DNA affects transfection efficiency.

▶ 03:48
3
Perform surgical injection and electroporation

Execute the surgical procedure to access the developing embryonic brain, inject plasmid DNA via capillary, and apply electrical pulses to drive transfection. This step requires precise timing and electrode positioning.

▶ 04:47
4
Apply cortical and deep-tissue electroporation

Perform targeted electroporation in cortical and subcortical regions to achieve differential gene expression patterns. This allows studying spatially distinct aspects of brain development.

▶ 07:03
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