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Delivery of the Cas9/sgRNA Ribonucleoprotein Complex in Immortalized and Primary Cells via Virus-like Particles (“Nanoblades”)
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Delivery of the Cas9/sgRNA Ribonucleoprotein Complex in Immortalized and Primary Cells via Virus-like Particles (“Nanoblades”)

Delivery of the Cas9/sgRNA Ribonucleoprotein Complex in Immortalized and Primary Cells via Virus-like Particles (“Nanoblades”)

DOI:
10.3791/62245-v

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09:23 min

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March 31, 2021

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, , ,
1LBMC, Laboratoire de Biologie et Modélisation de la Cellule Univ Lyon, ENS de Lyon, Université Claude Bernard Lyon 1, CNRS, UMR 5239, INSERM, U1210, Lyon, 69007, France, 2CIRI, Centre International de Recherche en Infectiologie Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon, F-69007, Lyon, France

Chapters

  • 00:05Introduction
  • 00:56Nanoblade Preparation
  • 02:45Concentration of Nanoblades on a Sucrose-Cushion
  • 04:16Monitoring Cas9 Loading Within Nanoblades and Transduction of Cells
  • 06:24Results: Characterization of Nanoblades
  • 08:29Conclusion

Summary

자동 번역

자동 번역

We have developed a simple and inexpensive protocol to load Cas9/single-guide RNA (sgRNA) ribonucleoprotein complexes within virus-like particles. These particles, called "Nanoblades", allow efficient delivery of the Cas9/sgRNA complex in immortalized and primary cells as well as in vivo.

Tags

Keywords: Cas9SgRNARibonucleoproteinVirus-like ParticlesNanobladesGenome EditingHEK 293T CellsTransfectionUltracentrifugationSucrose Gradient

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