In utero electroporation was performed as described previously (Ka and Kim 2018 (link); Ka et al. 2016a (link)). Briefly, timed pregnant female mice from E13.5 of gestation were deeply anesthetized, and the uterine horns were gently exposed by laparotomy. The lateral ventricles of an embryonic brain were injected with plasmid DNA (2 μg/μl) and 0.001% fast green (Sigma-Aldrich) using a Picospritzer II (Parker Hannifin). Electroporation was achieved by placing two sterile forceps-type electrodes on opposing sides of the uterine sac around the embryonic head and applying a series of short electrical pulses using a BTX ECM 830 electroporator (Harvard Apparatus) (five pulses with 100 ms length separated by 900 ms intervals were applied at 45 V).
Protocol detail
In utero Electroporation for Embryonic Brain
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Electrical
Electroporation
Embryonic
Fast green
Forceps
Gestation
Head
Laparotomy
Lateral ventricles of brain
Mice
Plasmid
Pregnant female
Pulses
Sterile
Uterine
Uterine horns
Corresponding Organization : Kent State University
Other organizations : Korea Institute of Toxicology, University of California, San Francisco
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Variable analysis
independent variables
- Injection of plasmid DNA (2 μg/μl) and 0.001% fast green (Sigma-Aldrich) into the lateral ventricles of an embryonic brain
- Application of a series of short electrical pulses using a BTX ECM 830 electroporator (Harvard Apparatus) (five pulses with 100 ms length separated by 900 ms intervals were applied at 45 V)
- Not explicitly mentioned
- Timed pregnant female mice from E13.5 of gestation
- Surgical procedure, including deeply anesthetizing the female mice and exposing the uterine horns by laparotomy
- Positive control: Not mentioned
- Negative control: Not mentioned
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