Using genome-wide CRISPR library screening with library resistant DCK to find new sources of Ara-C drug resistance in AML

• DOI: 10.1038/srep36199
• Publication Date: 2016-11-03T10:21:31Z
• Authors: Morito Kurata; Susan K. Rathe; Natashay J. Bailey; Natalie K. Aumann; Justine M. Jones; G. Willemijn Veld...; Branden S. Moriarity; David A. Largaespada
• Keywords: Deoxycytidine kinase

Abstract Acute myeloid leukemia (AML) can display de novo or acquired resistance to cytosine arabinoside (Ara-C), a primary component of induction chemotherapy. To identify genes capable independently imposing Ara-C resistance, we applied genome-wide CRISPR library human U937 cells and exposed them Ara-C. Interestingly, all drug resistant clones contained guide RNAs for DCK . avoid gene modification, gRNA cDNA was created by the introduction silent mutations. The screening repeated using , loss SLC29A identified as also being conveying resistance. determine if Dck results in increased sensitivity other drugs, conducted screen 446 FDA approved drugs two Dck-defective BXH-2 derived murine AML cell lines their sensitive parental lines. Both showed an increase prednisolone. Guide RNA rescue legitimate strategy multiple deficient were established with one clone becoming prednisolone sensitive. leukemic may become indicating be effective adjuvant therapy some cases DCK-negative AML.