In Utero Electroporation
For successful In Utero Electroporation (IUE) transfection efficiency, the delivery of a controlled electric current is one of key critical success factors.
As evident from the review by H Tabata and K Nakajima in their book ‘Electroporation and Sonoporation in Developmental Biology’, (Chapter 14, In Utero Electroporation: Assay System for Migration of Cerebral Cortical Neurons, p 143 – 152), the optimal current for most IUEs is 40-60mA.
For the following reasons:
– the electric impedance (resistance) for IUE is about 300-500 ohms in general
– but the impedance is not so stable because the application is in vivo
– and the output current does not exactly reflect Ohm’s law
We recommend that you set a 30-35V to achieve a 40-60mA output when using the CUY21SC Electroporation System.
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Electrode Recommendation:
With respect to in utero Electroporation, we recommend the following:
– CUY650P3 , CUY650P2 and CUY650P1 (If the mouse embryo is E10 – E12.5)
– CUY650P5 (If the mouse embryo is E13 or later)
With the CUY650P1 electrode, the transfection site could be smaller.
For information, the embryos are young (such as E10 or E11) light can be helpful to see inside the uterine wall. But it is important that the light temperature must not be hot.
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Protocol Recommendation:
Kindly note the following ‘In Utero Electroporation Resource’ file which includes the following articles:
– ‘Manipulating gene expressions by electroporation in the developing brain of mammalian embryos’
– ‘Multipolar Migration: The Third Mode of Radial Neuronal Migration in the Developing Cerebral Cortex’
– ‘Radial Migration of Superficial Layer Cortical Neurons Controlled by Novel Ig Cell Adhesion Molecule MDGA1′
– ‘Mammalian BarH1 Confers Commissural Neuron Identity on Dorsal Cells in the Spinal Cord’
– Technical Drawing of CUY651 electrode
– In utero Electroporation Parameters
– Technical Drawing of CUY650P1, P3 electrodes
– Technical Drawing of CUY650-5, P5, 7, P7 electrodes
– Photo of mouth-controlled micropipette system
Download: ‘In Utero Electroporation Resource’ file
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Kindly also note the following in utero Brain Applications. Identify the Application of interest and then click on the relevant Protocol to download it.
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The application titled above as ‘In Utero Electrodes’ identifies appropriate protocols for in utero electroporation.
For in utero electroporation parameters with Adult Mice Brain; please note the following links.
– ‘Shocking Development in Chick Embryology Electroporation and in ovo Gene Expression’
– ‘Gene Transfer into Cultured Mammalian Embryos by Electroporation’
Kindly also note the following link to Prof. Saito (Chiba University) explaining, in detail, best practise in utero electroporation procedure and protocol.