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HOX Loci fokusert CRISPR/sgRNA biblioteket Screening identifisere kritiske CTCF grenser
JoVE Journal
Genetics
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JoVE Journal Genetics
HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
DOI:

10:10 min

March 31, 2019

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Chapters

  • 00:04Title
  • 00:49The High Titer sgRNA Library Lentivirus Generation
  • 01:44Titration of Lentiviral Library in MOLM13 Leukemia Cells
  • 02:59Transduction of the Pooled CRISPR-Cas9 KO Library
  • 04:20Screening of the Pooled CRISPR-Cas9 KO Library with One-Step RT-qPCR
  • 05:41Verification of Integrated sgRNAs Positive Clones through Genotyping and Sanger Sequence
  • 06:37Detection of sgRNAs Induced Indel Mutation by Nuclease Digestion Assay
  • 07:48Results: Pooled CRISPR-Cas9 KO Library Screening and Identification
  • 09:19Conclusion

Summary

Automatic Translation

En CRISPR/sgRNA bibliotek er utlignet til avhør protein-koding gener. Muligheten for et sgRNA-bibliotek for å avdekke funksjonen til en CTCF grense i genet regulering imidlertid uutforsket. Her beskriver vi et HOX loci bestemte sgRNA bibliotek for å belyse funksjon CTCF grenser i HOX loci.

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