Mouse/Rat – Muscle

Electrode Recommendation:

There are three methods by which one can transfect gene into Mouse/Rat Muscle.

The first method involves injecting a pair of needle electrodes CUY560-5/-10 into the muscle above skin.  Since no surgery is involved, the researcher can easily perform electroporation and do so consecutively in a short period of time.  In our opinion, this is the best method.  However, as the volume of muscle affects the resistance value, that is, actual current value, the researcher needs to practice picking up the same volume of muscle by his/her fingers every time the experiment is performed.

Technical Drawing: CUY560-5 & CUY560-10,15

The second method requires one to first cut open the skin of the limb and then pick the muscle up with a tweezers type electrode CUY650P7/10 (refer to fig. a, P29 of attached article.  This is the method used by Dr. Takeshita in her paper.  She cuts and opens the leg, injects DNA buffer into the exposed muscle, grasps the injected muscle with a tweezers-type electrode (CUY650P5) and then applies square pulses.  If one observes the protocol; the impedance is from 218 ohms to 298 ohms.  When the impedance value is in this range, the current value will be stable and this will lead to the high reproducibility of the experiments.  The impedance is adjusted by changing the position of electrodes slightly before electroporation.

Technical Drawing: CUY650P5

The third method also requires one to first cut open the skin of the limb and then a pair of needle electrodes CUY560-5/-10 is injected directly into the muscle. If the one has no previous experience of electroporation with limb, we recommend this method.  Kindly note:

‘EP with rats limb – photo’

for photos of the relevant methodology/protocol.

Protocol Recommendation:

Kindly note the relevant information above and the following ‘Rat & Mouse Muscle EP Resource’ file.  This file includes the following articles:

-  ‘Elevated gastrin secretion by in vivo gene electroporation in skeletal muscle’

-  ‘Anti-monocyte chemoattractant protein-1 gene therapy attenuates pulmonary hypertension in rats’

-  ‘Gene Therapy for Central Diabetes Insipidus: Effective Antidiuresis by Muscle-Targeted Gene Transfer’

-  ‘Protection Against Autoimmune Myocarditis by Gene Transfer of Interleukin-10 by Electroporation’

-  ‘Skeletal muscle targeting in vivo electroporation-mediated HGF gene therapy of bleomycin-induced pulmonary fibrosis in mice’

-  ‘Gene Transfer into muscle by Electroporation in vivo’

-  ‘Toll-Like Receptor Adaptor Molecules Enhance DNA-Raised Adaptive Immune Responses against Influenza and Tumors through Activation of Innate Immunity’

Download ‘Rat & Mouse Muscle EP Resource’

Kindly also note the following EP Protocol for Gene Delivery into Mouse Leg at Yokohama City University:

* Reverse Polarity