Cell surface tag exchange (CSTE) system for tracking and manipulation of cells during integration of recombinase mediated cassette exchange of nucleic acid sequences into engineered receptor cells

摘要

A combinatorial system comprising two separate components, wherein the first component is a first cell surface tag (CST) exon flanked by a 3'intron fragment, and a tag encoding a gene of interest (GOI) in the antisense direction- Exchange donor vector (TEDV), and the second component encodes a second CST exon in which the entire intron sequence is contiguous to the exon encoding the transmembrane domain, and also a tag-exchange receptor site that encodes the reporter gene in the antisense direction ( TERS) in the genome, and the paired recombinase-mediated cassette exchange (RMCE) elements are contained in TEDV and TERS, and a reporter element for GOI encoded by TEDV when running RMCE between TEDV and TERS. Resulting in an exchange of, and an exchange of the first CST exon for the second CST exon, so that the derived engineered cell now expresses the first CST and GOI instead of the second CST and reporter gene.