Using RNA-seq and targeted nucleases to identify mechanisms of drug resistance in acute myeloid leukemia.

August 1, 2014
Authors: Rathe, et al.
B-MoGen Authors: Branden S Moriarity & David A Largaespada
Publication: Scientific Reports. 4: 6048. DOI:10.1038/srep0604
Published Date: August 2014
Characterizes new experimental partnership between deep-sequencing and CRISPR/TALEN technology for identifying the effects of gene changes on drug resistance.
Acute myeloid leukemia (AML) patients often become resistant to cytarabine (Ara-C), the primary component of induction chemotherapies. This study described the results from deep-sequencing of RNA derived from two Ara-C resistant AML) cell lines, and present CRISPR and TALEN based methods for accomplishing complete gene knockout (KO) in AML cells. They found protein-modifying loss-of-function mutations in DCK in both Ara-C resistant cell lines. CRISPR and TALEN-based KO of Dck dramatically increased the IC50 of Ara-C and introduction of a DCK overexpression vector into Dck KO clones resulted in a significant increase in Ara-C sensitivity. This effort demonstrates the power of using transcriptome analysis and CRISPR/TALEN-based KOs to identify and verify genes associated with drug resistance.