Nucleotide-resolution mapping of regulatory elements via allelic readout of tiled base editing - PubMed
5 hours ago
- #Immunotherapy
- #Gene Regulation
- #CRISPR
- Introduction of an end-to-end experimental assay and computational pipeline for nucleotide-resolution mapping of regulatory elements.
- Utilizes targeted sequencing of CRISPR-introduced alleles after dense base-editing mutagenesis.
- Proof of concept study on a CD19 enhancer identified crucial alleles and nucleotides for CD19 regulation.
- Visualization tools revealed transcription factor motifs corresponding to top-ranked nucleotides.
- Mutations in MYB, PAX5, and EBF1 binding sites were confirmed to reduce CD19 expression.
- Editing MYB and PAX5 motifs conferred resistance to CD19 CAR-T cell therapy, showing non-coding variants can drive immunotherapy escape.
- Approach achieves nucleotide-resolution genotype-phenotype mapping beyond conventional gRNA-based screens.