The inside scoop—evaluating gene delivery methods

AbstractHuman induced pluripotent stem cells (iPSCs) are established by introducing several reprogramming factors, such as OCT3/4, SOX2, KLF4, c-MYC. Because of their pluripotency and immortality, iPSCs are considered to be a powerful tool for regenerative medicine. To date, iPSCs have been established all over the world by various gene delivery methods. All methods induced high-quality iPSCs, but epigenetic analysis of abnormalities derived from differences in the gene delivery methods has not yet been performed. Here, we generated genetically matched human iPSCs from menstrual blood cells by using three kinds of vectors, i.e., retrovirus, Sendai virus, and episomal vectors, and compared genome-wide DNA methylation profiles among them. Although comparison of aberrant methylation revealed that iPSCs generated by Sendai virus vector have lowest number of aberrant methylation sites among the three vectors, the iPSCs generated by non-integrating methods did not show vector-specific aberrant methylation. However, the differences between the iPSC lines were determined to be the number of random aberrant hyper-methylated regions compared with embryonic stem cells. These random aberrant hyper-methylations might be a cause of the differences in the properties of each of the iPSC lines.

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Genetic modification of mesenchymal stem cells to enhance their anti-tumor efficacy

Asia-Pacific Journal of Oncology ◽

10.32948/ajo.2021.03.28 ◽

2021 ◽

pp. 1-6

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Gene Delivery ◽

Genetic Modification ◽

Chemokine Receptors ◽

Viral Vectors ◽

Viral Gene ◽

Delivery Methods ◽

Differentiation Potential ◽

Viral Gene Delivery

Non-hematopoietic mesenchymal stem cells (MSCs) are widely used in regenerative medicine and tissue engineering as they possess multilineage differentiation potential and self-renewal properties. MSCs can be easily isolated from several tissues and expanded following standard cell culture procedures. MSCs have the capability of mobilization to the tumor site; so, they can automatically relocate to the tumor sites through their chemokine receptors following intravenous transplantation. In this respect, they can be used for MSC-based gene therapy. In this therapeutic technique, beneficial genes are inserted by viral and non-viral methods into MSCs that lead to transgene expression in them. Genetic modifications of MSCs have been widely studied and thoroughly investigated to further enhance their therapeutic efficacy. The current strategies of MSC-based therapies emphasize the incorporation of beneficial genes, which will enhance the therapeutic ability of MSCs and have better homing efficiency. Non-viral methods produce less toxicity and immunogenicity compared to viral gene delivery methods and thus represent a promising and efficient tool for the genetic engineering of MSCs. Several non-viral gene delivery strategies have been developed in recent decades, and some of them have been used for MSCs modification. This mini review provides an overview of current gene delivery approaches used for the genetic modification of MSCs with beneficial genes including viral and non-viral vectors.

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