Chapter title |
Regulation of Receptor Tyrosine Kinases by miRNA: Overexpression of miRNA Using Lentiviral Inducible Expression Vectors.
|
---|---|
Chapter number | 13 |
Book title |
Receptor Tyrosine Kinases
|
Published in |
Methods in molecular biology, October 2014
|
DOI | 10.1007/978-1-4939-1789-1_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-1788-4, 978-1-4939-1789-1
|
Authors |
XiangDong Le, Andrew T Huang, Yunyun Chen, Stephen Y Lai, Le, XiangDong, Huang, Andrew T., Chen, Yunyun, Lai, Stephen Y., Andrew T. Huang, Stephen Y. Lai |
Abstract |
MicroRNAs have the ability to alter and regulate multiple genes, including RTK family members, making them an attractive approach for molecular therapeutic development. We use a pCDNA6.2-EmGFP-microRNA expression vector to overexpress individual mature microRNA and then transfer the expression cassette into a single, inducible lentiviral vector (pINDUCER20). We successfully use this system to create a pINDUCER-EmGFP-miRNA27a expression vector and generate a stable head and neck cancer cell line (UM-SCC-22A) that inducibly expresses miRNA-27a, resulting in targeted epidermal growth factor receptor down regulation. In this chapter, we describe the protocol for engineering the pINDUCER-EmGFP-microRNA expression vector, producing lentiviral particles for target cell infection, and evaluating downregulation of gene expression. |
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