How does Sendai virus reprogram cells?


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This video shows how the Sendai virus, present in the Cytotune®-iPS Sendai reprogramming kit, reprograms somatic cells to generate induced pluripotent stem cells (iPSCs)

How does the Sendai virus reprogram the cells?

Induced Pluripotent Stem Cells (iPSCs) are genetically reprogrammed somatic cells that exhibit a pluripotent stem cell state similar to that of embryonic stem cells. The discovery in 2006 that human and mouse fibroblasts could be reprogrammed to generate iPSCs with qualities remarkably similar to embryonic stem cells has created a new, valuable source of pluripotent cells.

Fibroblasts, similar to other somatic cell types, do not express high levels of Oct4, Sox2, Klf4 and c-Myc transcription factors under normal conditions. High levels of expression of these four genes will result in reprogramming of the fibroblast and become pluripotent.

There are several methods for generating iPSCs. Viruses such as retroviral vectors require the integration of the viral genome into the host chromosomes to express the reprogramming genes. Normal transcription of the gene allows the inserted Oct4, Sox2, Klf4 and c-Myc transgenes to be expressed with the host genes. The integration of viral DNA into the genome of the host disrupts the genome of the cells. This alteration may make iPSCs and their derivatives less safe for clinical application and may compromise compound screens or pathological pathway analyzes. On the other hand, Sendai virus is a negative-sense negative RNA virus that replicates in the cytoplasm. This means that it does not integrate into the genome of the host.

Sendai virus replicates independently of the cell cycle, unlike other approaches where exogenous genes are expressed only as cell divisions. Using this strategy, the Sendai virus produces a very large number of copies of the target gene. The Cytotune® iPS Sendai Reprogramming Kit contains four Sendai virus-based reprogramming vectors, each expressing one of the four factors Yamanaka Oct4, Sox2, Klf4 and c-Myc.

These viral vectors are added to the cell box to be reprogrammed, incubated overnight, and the reprogramming process begins. After reprogramming, quantitative PCR tests show that the Sendai virus does not remain in iPSCs, which allows you to perform your research with iPSCs that do not have genome integration or genomic integration. viral residues. Sendai virus is particularly useful when reprogramming blood cells derived from patients, such as CD34 positive cells, T cells and PBMCs.

Thanks to its non-integration capabilities, the Sendai virus from the Cytotune®-iPS Sendai Reprogramming Kit enables the use of iPSCs and their derivatives in a wide range of research experiments.



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