Investigating adverse genomic and regulatory changes caused by replacement of the full-length cDNA using Cas9 and AAV.

A "universal strategy" replacing the full-length cDNA may treat >99% of people with cystic fibrosis (pwCF), regardless of their specific mutations. Cas9-based gene editing was used to insert the cDNA and a truncated CD19 () enrichment tag at the locus in airway basal stem cells. This strategy restores CFTR function to non-CF levels.

Genomic patterns of transcription-replication interactions in mouse primary B cells.

Conflicts between transcription and replication machinery are a potent source of replication stress and genome instability; however, no technique currently exists to identify endogenous genomic locations prone to transcription-replication interactions. Here, we report a novel method to identify genomic loci prone to transcription-replication interactions termed transcription-replication immunoprecipitation on nascent DNA sequencing, TRIPn-Seq. TRIPn-Seq employs the sequential immunoprecipitation of RNA polymerase 2 phosphorylated at serine 5 (RNAP2s5) followed by enrichment of nascent DNA previously labeled with bromodeoxyuridine.