Transfection into Mouse/Rat Embryos In Utero by Electroporation

NEPA21

Tweezer Electrodes

Cables, Foot Switch & Others

Transfection into embryos: brain, spinal cord, inner ear and more.

APPLICATIONS
PUBLICATIONS

APPLICATIONS

Gene transfer into embryonic brains using in utero electroporation technique

1) Materials

2) In Utero Electroporation Procedure

Electroporation settings for ICR mice

Age	Electrode (diameter)	Voltage	Pulse On	Pulse Off	Number of Pulses
E12.5	3mm	33V	30msec	970ms	4
E13.5	5mm	30V	50msec	950ms	4
E14.5	5mm	33V	50msec	950ms	4
E15-	5mm	35V	50msec	950ms	4

If viability was prioritized over transfection efficiency, the number of pulses can be changed to two.

The actual current is displayed on the screen of the electroporator (NEPA21/CUY21/CUY21E, Nepa Gene). Make sure that the current would become 30-60mA. The current varies according to how the electrode applied or wetness of the uterine horn. Examine the gap between electrodes and the electrode contact areas to fit the current in the appropriate range. If the current is still above the range after the examination, change the voltage setting.

3) GFP expression

CAG-EGFP was injected into the both lateral ventricles of E14.5 mouse embryos and electronic pulses (33V, 50msec) were charged four times. 3 days later, the embryos (E17.5) were fixed and the brains were removed and examined under a fluorescence stereomicroscope (Fig. A).
Fluorescence was observed in the lateral region of the hemisphere onto which the anode had been placed and in the medial region of the opposite hemisphere.

Brains were frozen and sliced and the fluorescent image was obtained with a confocal laser microscope (Fig. B). GFP positive cells into which DNA was transferred at the ventricular zone (VZ) migrated to the intermediate zone (IZ) and cortical plate (CP). The arrowheads show the border between VZ and IZ and the border between IZ and CP. Dashed line show the border of tissues.
VZ: Ventricular Zone, IZ: Intermediate Zone, CP: Cortical Plate

Hidenori Tabata and Kazunori Nakajima, Department of Anatomy, Keio University School of Medicine

Transfection into Mouse/Rat Embryos In Utero by Electroporation

APPLICATIONS

Gene transfer into embryonic brains using in utero electroporation technique

1) Materials

2) In Utero Electroporation Procedure

3) GFP expression

PUBLICATIONS

Analyses of Conditional Knockout Mice for Pogz, a Gene Responsible for Neurodevelopmental Disorders in Excitatory and Inhibitory Neurons in the Brain

Lmo4 synergizes with Fezf2 to promote direct in vivo reprogramming of upper layer cortical neurons and cortical glia towards deep-layer neuron identities

Histological Analysis of a Mouse Model of the 22q11.2 Microdeletion Syndrome

p53/p21 pathway activation contributes to the ependymal fate decision downstream of GemC1

Variant-specific changes in RAC3 function disrupt corticogenesis in neurodevelopmental phenotypes

Tbr1 Misexpression Alters Neuronal Development in the Cerebral Cortex

Endosomal trafficking defects alter neural progenitor proliferation and cause microcephaly

Heterogeneous nuclear ribonucleoprotein U (HNRNPU) safeguards the developing mouse cortex

Impaired Function of PLEKHG2, a Rho-Guanine Nucleotide-Exchange Factor, Disrupts Corticogenesis in Neurodevelopmental Phenotypes

Simultaneous two-photon imaging of action potentials and subthreshold inputs in vivo

IgSF11 homophilic adhesion proteins promote layer-specific synaptic assembly of the cortical interneuron subtype

A pericellular hyaluronan matrix is required for the morphological maturation of cortical neurons

Huntington's disease alters human neurodevelopment

Knockdown of Son, a mouse homologue of the ZTTK syndrome gene, causes neuronal migration defects and dendritic spine abnormalities

DNA Repair Protein RAD51 Enhances the CRISPR/Cas9-mediated Knock-In Efficiency in Brain Neurons

In Vivo Imaging of the Coupling Between Neuronal and CREB Activity in the Mouse Brain

Prenatal Electroporation-Mediated Gene Transfer Restores Slc26a4 Knock-Out Mouse Hearing and Vestibular Function

Primary cilium-dependent cAMP/PKA signaling at the centrosome regulates neuronal migration

A Novel LGI1 Missense Mutation Causes Dysfunction in Cortical Neuronal Migration and Seizures

Role of Dach1 Revealed Using a Novel Inner Ear-Specific Dach1-knockdown Mouse Model

Estrogen Receptor α Promotes Cav1.2 Ubiquitination and Degradation in Neuronal Cells and in APP/PS1 Mice

In Vivo Single-Cell Genotyping of Mouse Cortical Neurons Transfected With CRISPR/Cas9

Both Excitatory and Inhibitory Neurons Transiently Form Clusters at the Outermost Region of the Developing Mammalian Cerebral Neocortex

Pathological mTOR Mutations Impact Cortical Development

Persistence of Learning-Induced Synapses Depends on Neurotrophic Priming of Glucocorticoid Receptors

Role of Per3, a Circadian Clock Gene, in Embryonic Development of Mouse Cerebral Cortex

MicroRNA-129-5p Is Regulated by Choline Availability and Controls EGF Receptor Synthesis and Neurogenesis in the Cerebral Cortex

Intersectional Monosynaptic Tracing for Dissecting Subtype-Specific Organization of GABAergic Interneuron Inputs

Single cell profiling of CRISPR/Cas9-induced OTX2 deficient retinas reveals fate switch from restricted progenitors

An Optogenetic Approach to Studies of the Mechanisms of Heterosynaptic Plasticity in Neocortical Neurons

Drebrin-like (Dbnl) Controls Neuronal Migration via Regulating N-Cadherin Expression in the Developing Cerebral Cortex

Comparative Evaluation of Genetically Encoded Voltage Indicators

Niche-derived laminin-511 Promotes Midbrain Dopaminergic Neuron Survival and Differentiation Through YAP

PLGF, a placental marker of fetal brain defects after in utero alcohol exposure.

Developmental activities of the complement pathway in migrating neurons.

PLGF, a placental marker of fetal brain defects after in utero alcohol exposure.

In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration.

Early postnatal GABAA receptor modulation reverses deficits in neuronal maturation in a conditional neurodevelopmental mouse model of DISC1.

Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions.

High-Throughput, High-Resolution Mapping of Protein Localization in Mammalian Brain by In Vivo Genome Editing.

Targeted in vivo genetic manipulation of the mouse or rat brain by in utero electroporation with a triple-electrode probe.

Early postnatal GABAA receptor modulation reverses deficits in neuronal maturation in a conditional neurodevelopmental mouse model of DISC1.

Radmis, a Novel Mitotic Spindle Protein that Functions in Cell Division of Neural Progenitors.

Screening for candidate genes involved in the production of mouse subventricular zone proliferative cells and an estimation of their changes in evolutionary pressure during primate evolution.

Development of cerebellar neurons and glias revealed by in utero electroporation: Golgi-like labeling of cerebellar neurons and glias.

Shootin1 Acts in Concert with KIF20B to Promote Polarization of Migrating Neurons.

Selective and regulated gene expression in murine Purkinje cells by in utero electroporation.

Persistent cortical plasticity by upregulation of chondroitin 6-sulfation.

Imaging brain electric signals with genetically targeted voltage-sensitive fluorescent proteins

Protein phosphatase 2A facilitates axonogenesis by dephosphorylating CRMP2.

Knockdown of DISC1 by in utero gene transfer disturbs postnatal dopaminergic maturation in the frontal cortex and leads to adult behavioral deficits.

Neurotrophin-3 is involved in the formation of apical dendritic bundles in cortical layer 2 of the rat.

Involvement of the cytoplasmic C-terminal domain of connexin43 in neuronal migration.

Electroporation-mediated gene transfer to the developing mouse inner ear.

Labeling embryonic mouse central nervous system cells by in utero electroporation.

Sirt1 contributes critically to the redox-dependent fate of neural progenitors

RECK modulates Notch signaling during cortical neurogenesis by regulating ADAM10 activity

Direct evidence that ventral forebrain cells migrate to the cortex and contribute to the generation of cortical myelinating oligodendrocytes.

Radial Migration of Superficial Layer Cortical Neurons Controlled by Novel Ig Cell Adhesion Molecule MDGA1

In vivo electroporation in the embryonic mouse central nervous system

G Protein betagamma Subunits and AGS3 Control Spindle Orientation and Asymmetric Cell Fate of Cerebral Cortical Progenitors

Birth-date-dependent segregation of the mouse cerebral cortical neurons in reaggregation cultures

Commissural neuron identity is specified by a homeodomain protein, Mbh1, that is directly downstream of Math1

Neuronal generation, migration, and differentiation in the mouse hippocampal primoridium as revealed by enhanced green fluorescent protein gene transfer by means of in utero electroporation

Hes genes regulate size, shape and histogenesis of the nervous system by control of the timing of neural stem cell differentiation

A NUDEL-dependent mechanism of neurofilament assembly regulates the integrity of CNS neurons

Generation of Reelin-Positive Marginal Zone Cells from the Caudomedial Wall of Telencephalic Vesicles

Layering defect in p35 deficiency is linked to improper neuronal-glial interaction in radial migration

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

The in vivo roles of STEF/Tiam1, Rac1 and JNK in cortical neuronal migration

Serine 732 Phosphorylation of FAK by Cdk5 Is Important for Microtubule Organization, Nuclear Movement, and Neuronal Migration

Mammalian BarH1 Confers Commissural Neuron Identity on Dorsal Cells in the Spinal Cord

Emx2 patterns the neocortex by regulating FGF positional signaling