Journal
/
Bioengineering
/
Electroporation-Mediated Delivery of Cas9 Ribonucleoproteins and mRNA into Freshly Isolated Primary Mouse Hepatocytes
JoVE Journal
Bioengineering
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Bioengineering
Electroporation-Mediated Delivery of Cas9 Ribonucleoproteins and mRNA into Freshly Isolated Primary Mouse Hepatocytes

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

DOI:
10.3791/63828-v

•

09:45 min

•

June 02, 2022

•

, , , ,
1Department of Bioengineering,Clemson University, 2Godley Snell Research Center,Clemson University

Chapters

  • 00:04Introduction
  • 01:18Animal Surgery
  • 03:26Hepatocyte Isolation
  • 04:54Electroporation and Cell Culture
  • 07:46Results: Generating Electroporation-Assisted CRISPR-Cas9 Edits in Freshly Isolated Mouse Hepatocytes
  • 09:06Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

ElectroporationCas9 RibonucleoproteinsMRNAPrimary Mouse HepatocytesGene EditingLiver PerfusionHepatocyte IsolationIn Vitro And In Vivo Disease ModelsAnesthesiaPortal VeinInferior Vena CavaPerfusion SolutionFlow RateLiver SofteningCotton SwabForceps

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Self-reporting Scaffolds for 3-Dimensional Cell Culture

Porous Silicon Microparticles for Delivery of siRNA Therapeutics

Delivery of Proteins, Peptides or Cell-impermeable Small Molecules into Live Cells by Incubation with the Endosomolytic Reagent dfTAT

Microfluidic Flow Chambers Using Reconstituted Blood to Model Hemostasis and Platelet Transfusion In Vitro

Manufacture and Drug Delivery Applications of Silk Nanoparticles

A Microfluidic Flow Chamber Model for Platelet Transfusion and Hemostasis Measures Platelet Deposition and Fibrin Formation in Real-time

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System

Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex

Handling and Assessment of Human Primary Prostate Organoid Culture

Encapsulated Cell Technology for the Delivery of Biologics to the Mouse Eye

Read Article