The β-globin Replicator greatly enhances the potential of S/MAR based episomal vectors for gene transfer into human haematopoietic progenitor cells.

ABSTRACT: Specific human chromosomal elements enhance the performance of episomal gene-transfer vectors. S/MAR-based episomal vector pEPI-eGFP transfects CD34⁺ haematopoietic cells, but only transiently. To address this issue we reinforced (1) transgene transcription by replacing the CMV promoter driving eGFP with the EF1/HTLV or SFFV promoters to produce vectors pEPI-EF1/HTLV and pEPI-SFFV, respectively; and (2) plasmid replication by inserting the replication-Initiation Region (IR) from the β-globin locus into vector pEPI-SFFV to produce vector pEP-IR. All vectors supported stable transfections in K562 cells. Transfections of CD34⁺ cells from peripheral blood of healthy donors reached 30% efficiency. Upon evaluation of CD34⁺/eGFP⁺ cells in colony-forming

SUBMITTER: Stavrou EF 

PROVIDER: S-EPMC5247744 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature