Tagmentation-based analysis reveals the clonal behavior of CAR-T cells in association with lentivector integration sites.

ABSTRACT: Integration site (IS) analysis is essential in ensuring safety and efficacy of gene therapies when integrating vectors are used. Although clinical trials of gene therapy are rapidly increasing, current methods have limited use in clinical settings because of their lengthy protocols. Here, we describe a novel genome-wide IS analysis method, "detection of the integration sites in a time-efficient manner, quantifying clonal size using tagmentation sequencing" (DIStinct-seq). In DIStinct-seq, a bead-linked Tn5 transposome is used, allowing the sequencing library to be prepared within a single day. We validated the quantification performance of DIStinct-seq for measuring clonal size with clones of known IS. Using ex vivo chimeric antigen receptor (CAR)-T cells, we revealed the characteri

SUBMITTER: Kim J 

PROVIDER: S-EPMC10285042 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature