Gene Delivery and Expression Systems in Induced Pluripotent Stem Cells

Abstract

Induced pluripotent stem (iPS) cells, which can be generated from somatic cells by genetic manipulation, are invaluable experimental and therapeutic tools for development of tissue regeneration technologies. Many studies have demonstrated that gene delivery to pluripotent stem cells is useful for basic studies in developmental biology and for driving differentiation toward a specifi c cell lineage for regenerative applications. Several gene delivery systems using viral and nonviral vectors have been used for stem cell research. These gene delivery systems are designed to accommodate specifi c research purposes; thus, each of them possesses its own advantages and disadvantages according to the experimental design. In addition, the type of constitutive promoter in the expression vector greatly affects the transcriptional activity of transgenes in pluripotent stem cells. Therefore, it is necessary to consider the characteristics of the vectors and their promoters when selecting a gene delivery system to transfer the target gene into iPS cells. In this mini-review, characteristics of commonly used viral (adenoviral, adeno-associated viral, retroviral, and lentiviral) vectors and a nonviral piggyBac transposon DNA vector with constitutive promoters are outlined to support the selection of an appropriate gene delivery and expression system for iPS cell research.

The use of induced pluripotent stem cells in domestic animals: a narrative review

“…Originally described in human models, these systems have since been applied to domestic species, including dogs [ 88 , 89 ], chickens [ 20 ], and cattle [ 152 ]. The wide range of reprogramming system options is beyond the scope of this article but have been reviewed elsewhere [ 153 – 155 ].…”

Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology

“…The addition of fluorescent reporters or antibiotic resistance cassettes can help to assess the transfection efficacy and allow the positive selection of transfected cells. Expression plasmids should carry suitable promoters to avoid transcriptional silencing that can occur in hPSCs [21]. Moreover, when using plasmid expression systems, DNA toxicity, potential unwanted insertion into the genome, and prolonged expression that can increase the chance of introducing undesired CRISPR-induced off-target (OT) mutations should be taken into account [22].…”

Efficient Modulation of TP53 Expression in Human Induced Pluripotent Stem Cells

“…Some promotors can be silenced over time or show no activity in stem cells (Chung et al, 2002;Xia, Zhang, Zieth, & Zhang, 2007). If cell transduction does not work, it would be best to try different promotors (e.g., EF1α; Zhang et al, 2017). At first, we used a cytomegalovirus promoter; however, after cloning the genes into an EF1α promoterdriven vector, we could transduce the hiPSCs.…”