Rhesus macaque autologous transplantation and hematopoietic barcoding. (A) Experimental summary. The replication-incompetent HIV-derived lentiviral barcoding vector used is diagrammed at the top right. The barcode consists of a 6–base pair library identification (ID) followed by a 35–base pair high-diversity cellular barcode. This vector was used to transduce rhesus macaque CD34⁺ cells, and these cells were reinfused after myeloablative total-body irradiation (TBI; 1000 rads) of the autologous recipient. Purified blood cells from various lineages underwent low-cycle polymerase chain reaction (PCR) amplification utilizing primers bracketing the barcode (red arrows in diagram), followed by Illumina sequencing and data processing, as described in supplemental Data. (B) Transplantation and engraftment parameters. The table summarizes CD34⁺ cell collection, transduction, and transplantation parameters, as well as total clone numbers and clone frequencies for each animal, after applying the threshold of a clone contributing at least 0.05% to at least 1 cell type at a minimum of at least 1 time point. GFP, green fluorescent protein; FACS, fluorescence-activated cell sorting; LTR, long terminal repeat; WPRE, woodchuck hepatitis posttranscriptional regulatory element.