Chemo- versus immuno-precipitation of G-quadruplex-DNA (G4DNA): a direct comparison of the efficiency of the antibody BG4 versus the small-molecule ligands TASQs for G4 affinity capture
Résumé
Abstract The search for genomic G-quadruplex (G4) motifs is motivated by their involvement in key cellular processes ( e.g ., transcription, replication) and corresponding dysregulations underlying genetic diseases. Sequencing-based methods have been developed to assess G4 DNA formation genome-wide, including G4-seq that detects G4s in purified DNA (so called in vitro ) from human lymphocytes using the G4 stabilizer PDS, and G4 ChIP-seq that detects G4s in the chromatin of human keratinocytes (so called in vivo ) using the G4-specific antibody BG4. We recently reported on the use of a small molecule, BioTASQ, as a proxy for BG4 for assessing the transcriptomic prevalence of G4-RNA in vivo using G4RP-seq. Here, we provide the first direct and unbiased comparison of the G4 capturing ability of small molecules BioTASQ and its new derivative BioCyTASQ versus the antibody BG4, developing G4DP-seq which makes the genome-wide detection of G4s in vitro and in vivo more straightforward, reproducible and reliable.
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