Genome-wide CRISPR/Cas9 library screening identified OGDH as a regulator of disease progress and HMAs-resistance in MDS by reprogramming glutamine metabolism

Resistance of hypomethylating agents (HMAs) remains a challenge in myelodysplastic neoplasms (MDS), leading to a poor prognosis. Altered metabolism is also reported to be closely associated with drug resistance in hematological malignancies. Here, we conducted a genome-wide CRISPR/Cas9 library screening to identify candidate genes involved in drug sensitivity. Overall design: Library: Use a genome-wide CRISPR/Cas9 knockout library (e.g., GeCKO v2.0) containing sgRNAs targeting all known genes. Lentiviral Packaging: Produce lentivirus containing the CRISPR library by co-transfecting HEK293T cells with the library and packaging plasmids.Experimental Design Using SKM-1 Cell Line for Decitabine Treatment and Sequencing Analysis at Day 0, Day 7, and Day 14. The NEBNext® Ultra™ DNA Library Prep Kit for Illumina® to prepare them for next-generation sequencing (NGS) on an Illumina HiSeq platform.