Abstract 1369: DNA, protein, and cell line reagents to facilitate RAS pathway drug discovery efforts

Abstract

Abstract The RAS Reagents Core within the NCI RAS Initiative produces and distributes qualified DNA vectors, cell lines, and protein production reagents to the external community to facilitate research into RAS biology and development of RAS cancer therapeutics. DNA vectors representing a variety of RAS mutations, as well as a complete collection of 180 genes in the RAS pathway are available as Gateway-compatible Entry clones. These clones represent the most commonly expressed transcript forms of the 180 genes, many of which were not previously available in full-length, wildtype forms. The vectors can be used with the FNLCR combinatorial cloning library to easily generate a large number of different types of expression clones with various promoters, fusion tags, and plasmid backbones. Cell line reagents include a variety of Ras-dependent mouse embryonic fibroblasts (MEFs) which have a defined mutant allele of KRAS and which serve as isogenic cell lines for investigating allele and mutant specific RAS biology, as well as for valuable screening tools. Protein reagents include E. coli and insect cell constructs and cell lines for production of a variety of Ras-related proteins. In particular, we have materials for the high yield production of properly processed, prenylated KRAS protein using a unique engineered insect cell expression system. This system can also be used to produce other prenylated small GTPases at levels not previously attainable. Many of the resources of the RAS Reference Core are readily available to the academic community through repositories or material transfer agreements, and can be licensed to industry scientists as well. The program is also interested in identifying the needs of the community for additional RAS-related resources, and is developing methods for vetting requests from the outside to provide additional reagents. Citation Format: Dominic Esposito. DNA, protein, and cell line reagents to facilitate RAS pathway drug discovery efforts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1369. doi:10.1158/1538-7445.AM2017-1369