Successfully transfected primary peripherally mobilized human CD34+ hematopoietic stem and progenitor cells (HSPCs) demonstrate increased susceptibility to retroviral infection

Abstract

Transfection, the process of introducing purified nucleic acids into cells, and viral transduction, viral-mediated nucleic acid transfer, are two commonly utilized techniques for gene delivery in the research setting. Transfection allows purified nucleic acid to be introduced into target cells through chemical-based techniques, nonchemical methods or particle-based methods, while viral transduction employs genomes or vectors based on adenoviruses, retroviruses (e.g. lentiviruses), adeno-associated viruses, or hybrid viruses. Transfected DNAs are often tested for potential effects on subsequent transduction, but it is not clear whether transfection itself rather than the particular nucleic acid being introduced might impact subsequent viral transfection. We observed a significant association between successfully transfected mobilized peripheral blood CD34+ human stem and progenitor cells (HSPCs) and permissiveness to subsequent lentiviral transduction, which was not evident in other cells such as 293 T cells and Jurkat cells. This association, apparently specific to CD34+ human stem and progenitor cells (HSPCs), is critical to both research and clinical applications as these cells are a frequent target of transfection and viral transduction owing to the durable nature of these cells in living systems. This finding may also present a significant opportunity to enhance the success of viral transduction for clinical applications.

Methods to Investigate miRNA Function: Focus on Platelet Reactivity

“…100 Of note, a recent study showed that prior transfection of CD34þ cells with plasmid DNA increases the efficiency of the following transduction procedures by threefold. 101 Stable regulation of gene expression has been reported in different studies using CRISPR/Cas9 tools 102 and Transcription Activator-Like Effector Nucleases (TALEN), 103 which are robust gene-editing systems to modify miRNA levels. In a recent study, over 80% of hPSCs were shown to undergo targeted genome editing using CRISPR-Cas9 reagents and nucleofection.…”

“… 100 Of note, a recent study showed that prior transfection of CD34+ cells with plasmid DNA increases the efficiency of the following transduction procedures by threefold. 101 …”