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电穿孔介导的Cas9核糖核蛋白和mRNA递送到新鲜分离的初级小鼠肝细胞中
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Electroporation-Mediated Delivery of Cas9 Ribonucleoproteins and mRNA into Freshly Isolated Primary Mouse Hepatocytes
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电穿孔介导的Cas9核糖核蛋白和mRNA递送到新鲜分离的初级小鼠肝细胞中

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

자동 번역

자동 번역

该协议描述了从肝脏中分离原代小鼠肝细胞并将CRISPR-Cas9电冲为核糖核蛋白和mRNA的技术，以破坏与肝脏遗传性代谢疾病相关的治疗靶基因。所描述的方法导致电穿孔后的高生存能力和高水平的基因修饰。

Tags

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

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