Next generation sequencing of antibody repertoires
Due to the size and duplication of the Ig loci, it has been impossible to quantify which V genes are used for recombination, and how, and why. We are using next generation sequencing assays (RNA-seq, ChIP-seq) to gain a high-resolution picture of underlying mechanisms, e.g. ncRNA transcription, at individual V genes and intergenic regions.
We have also developed an NGS assay (VDJ-seq), that can interrogate recombined immunoglobulin gene repertoires at unprecedented depth and resolution and provides the first unbiased and quantitative assay of V(D)J recombination, which will be a powerful resource for other researchers.