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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

Journal JoVE
Génétique

This content is Free Access.

Journal JoVE Génétique

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

•

09:16 min

•

September 01, 2019

•

Chi-Keung Cheng, Terry H.Y. Wong, Yuk-Lin Yung, Nelson C.N. Chan, Margaret H.L. Ng²

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

Chapitres

00:04Titre
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

Traduction automatique

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:RUNX1 Intronic Silencer CRISPR/Cas9 Ribonucleoprotein Acute Myeloid Leukemia Cis-regulatory Elements Gene Expression Transcriptional Role Hematopoietic Cells Electroporation Cas9 Nuclease GRNA RNP Complex Fragment Analysis

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