Summary
A major obstacle in studying monocyte cell biology has been the resistance of these cells to genetic manipulation, particularly when using nonviral methods. In the approach outlined in this chapter, we describe a strategy for stable gene silencing of monocytic cells. Using a vesicular stomatitis virus (VSV)-pseudotyped lentiviral vector expressing short hairpin RNA (shRNA), stable silencing of the p110α isoform of PI3-kinase in the human monocytic cell lines THP-1 and U-937 was achieved.
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Acknowledgments
The authors acknowledge Dr. Alice Mui for construction of the pHR-Gateway vector and for help with technical development. We also thank Dr. Greg Hannon for the gift of the pSHAG-1 vector. Work in the authors’ laboratory was supported by Canadian Institutes of Health Research (CIHR) operating grant MOP-8633 (NR).
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Lee, J.S., Reiner, N.E. (2009). Stable Lentiviral Vector-Mediated Gene Silencing in Human Monocytic Cell Lines. In: Reiner, N. (eds) Macrophages and Dendritic Cells. Methods in Molecular Biology™, vol 531. Humana Press. https://doi.org/10.1007/978-1-59745-396-7_18
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DOI: https://doi.org/10.1007/978-1-59745-396-7_18
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