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JoVE Journal Bioengineering
Electroporation-Mediated Delivery of Cas9 Ribonucleoproteins and mRNA into Freshly Isolated Primary Mouse Hepatocytes
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Electroporation-Mediated Delivery of Cas9 Ribonucleoproteins and mRNA into Freshly Isolated Primary Mouse Hepatocytes

Article DOI: 10.3791/63828-v • 09:45 min • June 2nd, 2022
June 2nd, 2022 •
*1, *1, 1, 2, 1
1Department of Bioengineering, Clemson University, 2Godley Snell Research Center, Clemson University
* These authors contributed equally

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Summary
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This protocol describes techniques for isolating primary mouse hepatocytes from the liver and electroporating CRISPR-Cas9 as ribonucleoproteins and mRNA to disrupt a therapeutic target gene associated with an inherited metabolic disease of the liver. The methods described result in high viability and high levels of gene modification after electroporation.

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Electroporation-mediated Delivery Cas9 Ribonucleoproteins MRNA Primary Mouse Hepatocytes Gene Editing In Vitro Disease Models In Vivo Disease Models Target Gene Knockdown Transplantation Of Gene-edited Hepatocytes Liver Perfusion Hepatocyte Isolation Cannulation Catheter Stability Troubleshooting Methods Tina Parker Ilayda Ates Callie Stuart Surgical Anesthesia U-shaped Lateral Incision Abdomen Skin Incision Peritoneum Cutting Organ Identification Portal Vein
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