KRAS.G12C Mutation and Effects on Proliferation in Human Lung Cancer

Abstract

A point mutation in GTPase KRAS is a known genetic driver of lung cancers. The current research aims to transform bacterial cells with a plasmid containing mutated KRAS and Hygromycin resistance cassette to be introduced into human lung cancer lines H1299 and A549 to ultimately produce stable expression of KRAS. PCR was used to amplify KRAS for cloning into hygromycin resistance cassette containing plasmid using restriction enzymes NheI and NotI. T4 DNA Ligase constructed plasmids then transformed into bacterial cells. Lipofectamine transfection agent introduced plasmids to H1299 and A549. Transient gene expression was confirmed by PCR, qPCR, and Western Blot.