Identifying therapeutic vulnerability in ataxia telangiectasia and Rad3-related protein (ATR) inhibitor resistant splicing factor mutant myeloid malignancies

The goal of this study is to undertake splicing analysis to validate clones derived from K562 cell lines which have been CRISPR engineered to carry mutations frequently observed in hematologic cancers on splicing factor U2af1. U2af1 specific guide RNAs which harbored specific mutations were used to target the nucleotides that code for Serine and Glutamine at amino acid positions 34 (S34) and 157 (Q157), respectively. Two different mutations each at U2af1S34 and U2af1Q157 loci were introduced individually in different K562 clones. While Serine at position 34 was converted to Phenylalanine or Tyrosine, Glutamine at position 157 was altered to Proline or Arginine. Subsequently, RNA-sequencing was carried out from individual clones.