Abstract

Genetic analysis in model organisms including yeast, C. elegans and fruit flies have yielded many fundamental discoveries in basic biology that ultimately resulted in breakthroughs in our understanding of the disease mechanisms of cancer. The lack of genetic tools in mammalian systems has significantly hampered genetics studies in human cells. The advent of RNAi technology has made it possible to conduct such analysis. 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 cells to create the desired genotype. 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. FY2014 ACCOMPLISHMENT. We have constructed cell lines with defined oncogenic perturbations, including KRAS mutations and we are currently analyzing the functional consequences of these alterations.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC011304-05
Application #
8938050
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2014
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