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O'Brien PJ, Elahipanah S, Rogozhnikov D, et al. Bio-Orthogonal Mediated Nucleic Acid Transfection of Cells via Cell Surface Engineering. ACS Cent Sci. 2017;3(5):489-500doi: 10.1021/acscentsci.7b00132.
O'Brien, P. J., Elahipanah, S., Rogozhnikov, D., & Yousaf, M. N. (2017). Bio-Orthogonal Mediated Nucleic Acid Transfection of Cells via Cell Surface Engineering. ACS central science, 3(5), 489-500. https://doi.org/10.1021/acscentsci.7b00132
O'Brien, Paul J, et al. "Bio-Orthogonal Mediated Nucleic Acid Transfection of Cells via Cell Surface Engineering." ACS central science vol. 3,5 (2017): 489-500. doi: https://doi.org/10.1021/acscentsci.7b00132
O'Brien PJ, Elahipanah S, Rogozhnikov D, Yousaf MN. Bio-Orthogonal Mediated Nucleic Acid Transfection of Cells via Cell Surface Engineering. ACS Cent Sci. 2017 May 24;3(5):489-500. doi: 10.1021/acscentsci.7b00132. Epub 2017 May 15. PMID: 28573212; PMCID: PMC5445537.
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ACS Cent Sci. 2017 May 24;3(5):489-500. doi: 10.1021/acscentsci.7b00132. Epub 2017 May 15.

Bio-Orthogonal Mediated Nucleic Acid Transfection of Cells via Cell Surface Engineering.

ACS central science

Paul J O'Brien, Sina Elahipanah, Dmitry Rogozhnikov, Muhammad N Yousaf

Affiliations

  1. Department of Chemistry and Biology, York University, Toronto, Canada, M3J 1P3.
  2. OrganoLinX Inc., Toronto, Canada.

PMID: 28573212 PMCID: PMC5445537 DOI: 10.1021/acscentsci.7b00132

Abstract

The efficient delivery of foreign nucleic acids (transfection) into cells is a critical tool for fundamental biomedical research and a pillar of several biotechnology industries. There are currently three main strategies for transfection including reagent, instrument, and viral based methods. Each technology has significantly advanced cell transfection; however, reagent based methods have captured the majority of the transfection market due to their relatively low cost and ease of use. This general method relies on the efficient packaging of a reagent with nucleic acids to form a stable complex that is subsequently associated and delivered to cells via nonspecific electrostatic targeting. Reagent transfection methods generally use various polyamine cationic type molecules to condense with negatively charged nucleic acids into a highly positively charged complex, which is subsequently delivered to negatively charged cells in culture for association, internalization, release, and expression. Although this appears to be a straightforward procedure, there are several major issues including toxicity, low efficiency, sorting of viable transfected from nontransfected cells, and limited scope of transfectable cell types. Herein, we report a new strategy (SnapFect) for nucleic acid transfection to cells that does not rely on electrostatic interactions but instead uses an integrated approach combining bio-orthogonal liposome fusion, click chemistry, and cell surface engineering. We show that a target cell population is rapidly and efficiently engineered to present a bio-orthogonal functional group on its cell surface through nanoparticle liposome delivery and fusion. A complementary bio-orthogonal nucleic acid complex is then formed and delivered to which chemoselective click chemistry induced transfection occurs to the primed cell. This new strategy requires minimal time, steps, and reagents and leads to superior transfection results for a broad range of cell types. Moreover the transfection is efficient with high cell viability and does not require a postsorting step to separate transfected from nontransfected cells in the cell population. We also show for the first time a precision transfection strategy where a single cell type in a coculture is target transfected via bio-orthogonal click chemistry.

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