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 (If the mouse embryo is younger than E12.5)

- CUY650P5 (If the mouse embryo is older than E12.5)

Kindly note below for the Technical Drawings of these electrodes.

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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.

Application	Protocol
CdK5 Kinase – Cortex (2003)	Serine 732 Phosphorylation of FAK by Cdk5 Is Important for Microtubule Organization, Nuclear Movement, and Neuronal Migration’
Central Nervous System (2006)	In Vivo Electroporation in the Embryonic Mouse Central Nervous System
Cerebral Cortical Nueron (2005)	Birth-Date-Dependent Segreation of the Mouse Cerebral Cortical Neurons in Re-Aggregation Cultures
Cerebral Cortical Progenitors (2005)	G Protein By Subunits and AGS3 Control Spindle Orientation and Asymmetric Cell Fate of Cerebral Cortical Progenitors
Cortex (2003)	Multipolar Migration: The Third Mode of Radial Neuronal Migration in the Developing Cerebral Cortex
Cortex – Brain (2001)	Efficient In Utero Gene Transfer System to the Developing Mouse Brain using Electroporation: Visulization of Neuronal Migration in the Developing Cortex
Cortex – Brain (2003)	Emx2 Patterns the Neocortex by Regulating FGF Positional Signalling
Cortical Neurons (2006)	Radial Migration of Superficial Layer Cortical Neurons Controlled by Novel Ig Cell Adhesion Molecule MDGA1
Glia Cortex – Brain (2003)	Layering Defect in p35 Deficiency is linked to Improper Neuronal-Glial Interaction in Radial Migration
Hes Gene – Telencephalon (2004)	Hes genes regulate size, shape and histogenesis of the nervous system by control of the timing of neural stem cell differentiation
Hippocampus (2005)	Neuronal Generation, Migration, and Differentation in the Mouse Hippocampal Primoridium as Revealed by Enhanced Green Fluorescent Protein Gene Transfer by Means of In Utero Electroporation
In Utero Electrodes	Electroporation Protocol; Technical Drawing of CUY651, CUY650P1, P3, CUY650-5, P5, 7, P7 Electrodes and Mouth-controlled Micropipette System Photo
JNK – Cerebral Cortex (2003)	The In Vivo Roles of STEF/Tiam1, Rac1 and JNK in Cortical Neuronal Migration
Neural Stem Cell – Brain (2001)	Roles of the Basic Helix-Loop-Helix Genes Hes1 and Hes5 in Expansion of Neural Stem Cells of the Developing Brain*
NUDEL – Brain (2004)	A NUDEL – Dependent Mechanism of Neurofilament Assembly Regulates the Integrity of CNS Neurons
Spinal Cord (2003)	Mammalian BarH1 Confers Commissural Neuron Identity on Dorsal Cells in the Spinal Cord
Spinal Cord (2005)	Commissural Neuron Identity is Specified by a Homeodomain Protein, Mbh1, that is Directly Downstream of Math1
Telencephalon (2004)	Generation of Reelin-Positive Marginal Zone Cells from the Caudomedial Wall of Telencephalic Vesicles

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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.

- ‘Locus-Specific Rescue of GluR1€ NMDA Receptors in Mutant Mice Identifies the Brain Regions Important for Morphine Tolerance and Dependence’

- ‘Multipolar Migration: The Third Mode of Radial Neuronal Migration in the Developing Cerebral Cortex’

- ‘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.