Abstract

PURPOSE.
We aim to develop new mammalian vector systems that express shRNAs and cDNAs efficiently. This will enable us to rapidly introduce defined genetic changes into normal cells to create the desired phenotypes. SIGNIFICANT MATERIALS AND METHODS. Expression vectors for shRNAs and cDNAs that enable the rapid immortalization of primary human cells and the introduction of multiple oncogenic lesions. FY2015 ACCOMPLISHMENT. We have constructed human small airway epithelial cell lines with defined oncogenic perturbations, including KRAS mutations and p53 loss, we showed that these alterations can lead to oncogenic transformation of these cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC011304-06
Application #
9153854
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2015
Total Cost
Indirect Cost

  Institution

Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code

  Publications

Brown, Zachary J; Fu, Qiong; Ma, Chi et al. (2018) Carnitine palmitoyltransferase gene upregulation by linoleic acid induces CD4+ T cell apoptosis promoting HCC development. Cell Death Dis 9:620
Sissung, Tristan M; Ley, Ariel M; Strope, Jonathan D et al. (2017) Differential Expression of OATP1B3 Mediates Unconjugated Testosterone Influx. Mol Cancer Res 15:1096-1105
Read, Abigail; Gao, Shaojian; Batchelor, Eric et al. (2017) Flexible CRISPR library construction using parallel oligonucleotide retrieval. Nucleic Acids Res :
Yuen, Garmen; Khan, Fehad J; Gao, Shaojian et al. (2017) CRISPR/Cas9-mediated gene knockout is insensitive to target copy number but is dependent on guide RNA potency and Cas9/sgRNA threshold expression level. Nucleic Acids Res 45:12039-12053
Porter, Joshua R; Fisher, Brian E; Baranello, Laura et al. (2017) Global Inhibition with Specific Activation: How p53 and MYC Redistribute the Transcriptome in the DNA Double-Strand Break Response. Mol Cell 67:1013-1025.e9
Smith, Jordan L; Lee, Liam C; Read, Abigail et al. (2016) One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation. Cell Biosci 6:57
Luo, Ji (2016) CRISPR/Cas9: From Genome Engineering to Cancer Drug Discovery. Trends Cancer 2:313-324
Weng, Meng-Tzu; Lee, Jih-Hsiang; Wei, Shu-Chen et al. (2012) Evolutionarily conserved protein ERH controls CENP-E mRNA splicing and is required for the survival of KRAS mutant cancer cells. Proc Natl Acad Sci U S A 109:E3659-67
Luo, Ji; Emanuele, Michael J; Li, Danan et al. (2009) A genome-wide RNAi screen identifies multiple synthetic lethal interactions with the Ras oncogene. Cell 137:835-48