Genetic Modification of Chondrocytes Using Viral Vectors

2010 ◽  
pp. 99-114 ◽  
Author(s):  
Teresa Coughlan ◽  
Aileen Crawford ◽  
Paul Hatton ◽  
Michael Barker
Keyword(s):  
Genetic Modification ◽  
Viral Vectors

scholarly journals Pseudotyping Lentiviral Vectors: When the Clothes Make the Virus

Viruses ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1311 ◽  
Author(s):  
Alexis Duvergé ◽  
Matteo Negroni
Keyword(s):  
Gene Therapy ◽  
Genetic Modification ◽  
Viral Vectors ◽  
Molecular Level ◽  
Lentiviral Vectors ◽  
Human Cells ◽  
Surface Proteins ◽  
Holy Grail ◽  
Viral Therapy

Delivering transgenes to human cells through transduction with viral vectors constitutes one of the most encouraging approaches in gene therapy. Lentivirus-derived vectors are among the most promising vectors for these approaches. When the genetic modification of the cell must be performed in vivo, efficient specific transduction of the cell targets of the therapy in the absence of off-targeting constitutes the Holy Grail of gene therapy. For viral therapy, this is largely determined by the characteristics of the surface proteins carried by the vector. In this regard, an important property of lentiviral vectors is the possibility of being pseudotyped by envelopes of other viruses, widening the panel of proteins with which they can be armed. Here, we discuss how this is achieved at the molecular level and what the properties and the potentialities of the different envelope proteins that can be used for pseudotyping these vectors are.

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

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.

scholarly journals An efficient electroporation protocol for the genetic modification of mammalian cells

2016 ◽  
Author(s):  
Leonardo Chicaybam ◽  
Camila Barcelos ◽  
Barbara Peixoto ◽  
Mayra Carneiro ◽  
Cintia Gomez Limia ◽  
...  
Keyword(s):  
Cell Lines ◽  
Genetic Modification ◽  
Mammalian Cells ◽  
Transgene Expression ◽  
Viral Vectors ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Sleeping Beauty ◽  
Primary Cells ◽  
Peripheral Blood Mononuclear

AbstractGenetic modification of cell lines and primary cells is an expensive and cumbersome approach, often involving the use of viral vectors. Electroporation using square wave generating devices, like Lonza´s Nucleofector, is a widely used option, but the costs associated with the acquisition of electroporation kits and the transient transgene expression might hamper the utility of this methodology. In the present work we show that our in house developed buffers, termed Chicabuffers, can be efficiently used to electroporate cell lines and primary cells from murine and human origin. Using the Nucleofector II device, we electroporated 14 different cell lines and also primary cells, like mesenchymal stem cells and cord blood CD34+, providing optimized protocols for each of them. Moreover, when combined with Sleeping Beauty based transposon system, long-term transgene expression could be achieved in all types of cells tested. Transgene expression was stable and did not interfere with CD34+ differentiation to committed progenitors. We also show that these buffers can be used in CRISPR-mediated editing of PDCD1 gene locus in 293T and human peripheral blood mononuclear cells. The optimized protocols reported in this study provide a suitable and cost-effective platform for the genetic modification of cells, facilitating the widespread adoption of this technology.

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