Project description:We deleted SLC2A5 using CRISPR-Cas9 technology in human lung cancer cell A549. Control A549 cells and A549 cells with SLC2A5 knockout were transplanted in balb/c nude mice. Then RNA-seq was performed in control A549 and SLC2A5 ablation A549 Xenografts.
Project description:To identify genes for radioresistance in lung cancer, unbiased CRISPR/Cas9 library knockout screening was performed by constructing stable cells in A549 containing a pooled genome-wide human sgRNA library containing 12, 3411 sgRNAs targeting 19,050 unique human genes. Two populations of cells were either nonirradiated or irradiated at 4 Gy, and they were cultured in vitro for 4 days. Then genomic DNA was acquired to readout the sgRNA representation by deep sequencing.
Project description:The widespread clinical application of paclitaxel (PTX) in cancer treatment has been significantly limited by the emergence of drug resistance and drug-tolerant persisters. Through genome-wide CRISPR/Cas9 screening, we identified cell division cycle 6 (CDC6) as a critical determinant of cell adhesion-mediated paclitaxel resistance. CDC6, an essential DNA replication licensing factor, functions through a pathway distinct from well-characterized resistance mechanisms. The depletion of CDC6 considerably increases paclitaxel-induced cell death. Beyond its established role in stabilizing chromosomes, our findings indicate that tropomodulin-3 (Tmod3) interacts with CDC6 in the cytoplasm. This interaction enhances CDC6 protein stability and drives drug-resistant phenotypes through regulating actin cytoskeleton remodeling and facilitating focal adhesion assembly. In addition, pharmacological or genetic inhibition of CDC6 notably sensitizes the antitumor efficacy of paclitaxel both in vitro and in vivo. Collectively, our studies elucidate the mechanisms through which CDC6 functions as a key regulator of paclitaxel resistance and provide a potential strategy to enhance paclitaxel efficacy through cytoskeletal-adhesion axis modulation.
Project description:Genome-wide CRISPR-Cas9 knockout screen using TKOv1 sgRNA library performed in isogenic RBM10-proficient and RBM10-deficient HCC827 cells.
Project description:Genome-wide CRISPR-Cas9 knockout screen using TKOv1 sgRNA library was performed in isogenic RBM10-proficient and RBM10-deficient HCC827 cells.
