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CRISPR/Cas9核糖核酸蛋白在急性骨髓性白血病细胞中的RUNX1电子消光器的转录作用研究
JoVE Journal
Genetics
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JoVE Journal Genetics
Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells
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CRISPR/Cas9核糖核酸蛋白在急性骨髓性白血病细胞中的RUNX1电子消光器的转录作用研究

DOI:
10.3791/60130-v

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09:16 min

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September 01, 2019

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, , , ,
1Blood Cancer Cytogenetics and Genomics Laboratory, Department of Anatomical and Cellular Pathology, Prince of Wales Hospital,The Chinese University of Hong Kong, 2State Key Laboratory in Oncology in South China,The Chinese University of Hong Kong

Chapters

  • 00:04Title
  • 01:16crRNA Design and Preparation of Cas9/gRNA RNP Complexes
  • 02:16Electroporation of RNP Complexes into OCI-AML3 Cells
  • 03:12Screening and Selection of Cell Clones with Biallelic Deletions
  • 05:51Functional Analyses of Silencer Deletions with Real-time Quantitative RT-PCR
  • 07:18Results: RUNX1 Intronic Silencer Deletion
  • 08:16Conclusion

Summary

Automatic Translation

Automatic Translation

直接交付预组装的Cas9/指南RNA核糖核蛋白复合物是造血细胞基因组编辑的快速而有效的方法。在这里，我们利用这种方法删除RUNX1电子消音器，并检查OCI-AML3白血病细胞的转录反应。

Tags

Keyword Extraction:RUNX1Intronic SilencerCRISPR/Cas9 RibonucleoproteinAcute Myeloid LeukemiaCis-regulatory ElementsGene ExpressionTranscriptional RoleHematopoietic CellsElectroporationCas9 NucleaseGRNARNP ComplexFragment Analysis

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