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One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster
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Neuroscienze
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JoVE Journal Neuroscienze
One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

DOI:
10.3791/64729-v

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07:23 min

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January 26, 2024

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, ,
1Department of Cell and Developmental Biology,University of Michigan, 2Cellular and Molecular Biology Program,University of Michigan, 3Michigan Neuroscience Institute Affiliate,University of Michigan

Capitoli

  • 00:00Introduction
  • 00:30Design and Construction of Gene Editing Reagents
  • 06:51Results: Analyzing PERIOD-mNeonGreen Protein Expression in Clock Neurons
  • 07:10Conclusion

Summary

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Here, we present a simplified endogenous gene tagging protocol for Drosophila, which utilizes a PCR-based technique for marker-free identification of successful genetic modifications, facilitating the development of stable knock-in lines.

Tags

Keywords: CRISPRGene TaggingDrosophila MelanogasterHomologous RecombinationFluorescent ProteinKnock-inSgRNADonor VectorCloningPlasmidSanger Sequencing

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