Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome

By N. Fusaki, H. Ban, Akiyo Nishiyama, Koichi Saeki, M. Hasegawa. 2009

1,410 citations recorded by the source corpus.

Induced pluripotent stem cells (iPSC) have been generated from somatic cells by introducing reprogramming factors. Integration of foreign genes into the host genome is a technical hurdle for the clinical application. Here, we show that Sendai virus (SeV), an RNA virus and carries no risk of altering host genome, is an efficient solution for generating safe iPSC. Sendai-viral human iPSC expressed pluripotency genes, showed demethylation characteristic of reprogrammed cells. SeV-derived transgenes were decreased during cell division. Moreover, viruses were able to be easily removed by antibody-mediated negative selection utilizing cell surface marker HN that is expressed on SeV-infected cells. Viral-free iPSC differentiated to mature cells of the three embryonic germ layers in vivo and in vitro including beating cardiomyocytes, neurons, bone and pancreatic cells. Our data demonstrated that highly-efficient, non-integrating SeV-based vector system provides a critical solution for reprogramming somatic cells and will accelerate the clinical application.

Subfields: Molecular Biology

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Citations in the corpus

Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors (2011, methodology): “However, to apply SeV vector technology to the generation of safer iPSCs, the issue of the sustained cytoplasmic replication of viral vectors after the iPSCs have been established had to be overcome, even though viral vectors are slowly diluted during the robust cell division of iPSCs and SeV vector”
Induced pluripotent stem cells: past, present, and future. (2012, neutral): “, 2008), Sendai virus (Fusaki et al., 2009), adenovirus (Stadtfeld et al.”
Induced pluripotency: history, mechanisms, and applications. (2010, background): “Human fibroblasts have also been reprogrammed into iPSCs with adenoviral vectors (Zhou and Freed 2009) and Sendai virus (Fusaki et al. 2009), as well as with polycistronic minicircle vectors (Jia et al. 2010) and self-replicating selectable episomes (Yu et al. 2009), albeit the latter system…”
Human iPSC-based Modeling of Late-Onset Disease via Progerin-induced Aging (2013, positive): “iPSC Derivation and Differentiation Apparently healthy young donor, old donor, and HGPS patient and Parkinson’s patient fibroblasts purchased from Coriell were reprogrammed as described (Fusaki et al., 2009) using CytoTune Sendai viruses (Life Technologies) at an MOI of ten.”
Gene therapy clinical trials worldwide to 2012 – an update (2013, background): “Originally produced by retroviral-mediated delivery, refinements to the system using non-integrating vectors and transient expression systems [77–81] will also address safety”
Highly efficient reprogramming to pluripotency and directed differentiation of human cells using synthetic modified mRNA (2010, methodology): “…derived with two DNA-free methods: serial protein transduction with recombinant proteins incorporating cell-penetrating peptide moieties (Kim et al., 2009; Zhou et al., 2009) and transgene . delivery using the Sendai virus, which has a completely RNAbased reproductive cycle (Fusaki et al., 2009).”