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利用 CRISPR/Cas9 Ribonucleoproteins 快速简便地生成线虫基因组点突变体的管道
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Genetics
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JoVE Journal Genetics
A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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利用 CRISPR/Cas9 Ribonucleoproteins 快速简便地生成线虫基因组点突变体的管道

DOI:
10.3791/57518-v

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08:37 min

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April 30, 2018

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, , , ,
1Department of Pharmacology,University of Michigan

Chapters

  • 00:04Title
  • 00:42sgRNA Target Selection and Homology-directed Repair Template Design
  • 02:21Prepare Injection Mix and Injection Protocol
  • 03:18Screen P0 Plates and Single mCherry(+) F1s
  • 04:03Single Worm PCR and Genotyping
  • 05:41Indentification and Sequence Verification of Edited Animals
  • 06:34Results: Rapid CRISPR/Cas9 Generated Point Mutations in C. elegans sod-1
  • 07:50Conclusion

Summary

Automatic Translation

Automatic Translation

在这里, 我们提出了一个方法, 以工程的基因组的C. 线虫使用 CRISPR-Cas9 ribonucleoproteins 和同源依赖修复模板。

Tags

CRISPR/Cas9Genomic Point MutationC. ElegansHomology Directed RepairSsODNSgRNARestriction Endonuclease SiteNeuroscienceNeurodegenerative DiseaseEndogenous Regulatory ControlProtein OverexpressionPipelineRapidFacile

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