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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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遗传学
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JoVE 杂志 遗传学
Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

DOI:
10.3791/59086-v

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07:56 min

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April 11, 2019

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, , ,
1Genome Institute of Singapore,Agency for Science Technology and Research, 2School of Chemical and Biomedical Engineering,Nanyang Technological University

Chapters

  • 00:04Title
  • 00:18Cell Passaging
  • 01:08Cell Transfection
  • 01:47Transfected Cell Fluorescence Activated Cell Sorting (FACS)
  • 02:32Individual Clone Expansion and Editing Efficiency Evaluation
  • 06:09Results: Representative Gene Editing Efficiency Analyses
  • 07:24Conclusion

Summary

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CRISPR-Cas is a powerful technology to engineer the complex genomes of plants and animals. Here, we detail a protocol to efficiently edit the human genome using different Cas endonucleases. We highlight important considerations and design parameters to optimize editing efficiency.

Tags

CRISPR-Cas9Genome EditingMammalian Cell LinesKnock-outKnock-inCell PassagingTrypsin-EDTATransfectionFACSFluorescent MarkerCell Culture MediumGenomic DNA Extraction

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