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