Abstract

Genetic analysis in model organisms such as yeast, C. elegans and fruit flies have resulted in many fundamental discoveries in basic biology. The lack of genetic tools in mammalian system has significantly hampered genetics studies in mammalian cells. The advent of RNAi technology have now made it possible to conduct such analysis. PURPOSE We aim to develop new mammalian vector systems to express shRNAs and cDNAs efficiently. This will enable us to introduce multiple, defined genetic changes into cells to create the desired genotype. SIGNIFICANT MATERIALS AND METHODS Expression vectors for shRNAs and cDNAs. FY2011 ACCOMPLISHMENT We have established a design for the vector and validated shRNAs against tumor suppressors.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC011304-02
Application #
8349457
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
2011
Total Cost
$141,392
Indirect Cost

  Institution

Name
National Cancer Institute Division of 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