Misexpression of the gene of interest by in ovo electroporation

NEPA21 / CUY21 Applications

[In Ovo] Misexpression of the gene of interest by in ovo electroporation

.

(1) Injection of the plasmid solution colored by Fast Green into the central canal

(2) A pair of electrodes are put on the vitelline membrane overlying the embryos, and a 25V, 50 ms rectangular pulse was charged 4 times.

(3) We can monitor the transfection efficiency by electroporating GFP expression vector.

.

.

(4) Strong GFP fluorescence could be detected even 48 hours after electroporation.

(5) Right side of the embryo is transfected, hence the left side could be served as the controll (Dorsal view)

.

.

Mechanisms of brain regionalization and neural circuit formation have been studied by misexpression of transcription factors (En1/2, Pax2/5/6, Otx2, Gbx2), secreted factors (Fgf, Shh, semaphoring), signal transduction molecule (Ras, Sprouty2) and receptors (neuropilin)

.

Graduate School of Life Sciences, and Institute of Development, Aging and Cancer, Tohoku University

Harukazu Nakamura and Yuji Watanabe

*Development Growth & Differentiation, Volume 42, Issue 3, Page 199-201, June 2000

.