Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9

13.3K Views

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08:27 min

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April 10th, 2018

DOI :

10.3791/57278-v

April 10th, 2018

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Lorenzo Brunetti*¹^,²^,³, Michael C. Gundry*¹^,²^,⁴, Ayumi Kitano⁴, Daisuke Nakada¹^,²^,⁴, Margaret A. Goodell¹^,²^,⁴^,⁵

¹Stem Cells & Regenerative Medicine Center, Baylor College of Medicine, ²Center for Cell and Gene Therapy, Baylor College of Medicine, ³Centro di Ricerca Emato-Oncologica (CREO), University of Perugia, ⁴Department of Molecular & Human Genetics, Baylor College of Medicine, ⁵Texas Children's Hospital & Houston Methodist Hospital

Summary

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Keywords CRISPR Cas9

Gene Disruption

Hematopoietic Progenitor Cells

Normal And Malignant Hematopoiesis

Primary T cells

Hematopoietic Cell Lines

Primary Leukemia Samples

Chapters in this video

0:05

Title

1:05

sgRNA Fwd Primer Design, DNA Template Synthesis, and ln Vitro Transcription of sgRNA

3:38

Cas9-sgRNA Complexing and Electroporation

5:45

Results: Use of Cas9-sgRNA Electroporation Protocol to Study Efficient Knockout of Target Gene

7:48

Conclusion

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