Abstract

PURPOSE.
We aim to develop better reagents, vector systems and methodologies to enable efficient gene silencing, gene activation, and gene editing in cancer cells. This will enable us to rapidly introduce defined genetic changes into cells to create the desired genotype and study the resulting phenotype. This will also enable us to carry out gain and loss of function genetic screens in cancer cells for target discovery. SIGNIFICANT MATERIALS AND METHODS. We have generated expression vectors for shRNAs, cDNAs and sgRNAs. We have also developed methods for the design and construction of CRISPR/Cas9 libraries. ACCOMPLISHMENT. We have developed reagents and methods for combinatorial gene knockdown and for targeted gene knockout in cancer cells. We have also developed CRISPR/Cas9 libraries that can be used for gene knockout screening in cancer cell lines.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC011304-08
Application #
9556518
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
8
Fiscal Year
2017
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