A facile methodology using quantum dot multiplex labels for tracking co-transfection

Jessica A. Kretzmann, Rui Lu Feng, Alaa M. Munshi, Diwei Ho, Anna M. Ranieri, Massimiliano Massi, Martin Saunders, Marck Norret, Cameron W. Evans, K. Swaminathan Iyer

Research output: Contribution to journalArticle

Abstract

Advances in the field of genome engineering demand the development of efficient non-viral transfection agents capable of delivering multiple distinct nucleic acids efficiently to cells (co-transfection). However, current delivery methods result in lower co-transfection efficiency than single plasmid transfections, and the efficiency decreases further with increasing numbers of plasmids. The development of a high-throughput methodology is required for the validation of co-transfection platforms to facilitate independent tracking of not only the multiple DNA plasmids during transfection but also the localisation of transfection agents. This is pivotal to determine the bottlenecks in achieving high transfection efficiencies at various stages of the cell internalisation and plasmid expression process. Herein we demonstrate that this can be achieved using a facile methodology in which quantum dots (QDs) are used to label two different plasmid DNA assemblies that are delivered to cells simultaneously using a dendronised polymer system. Multispectral confocal imaging can be used to separate signals from each polyplex as well as the expressed fluorescent reporter proteins to determine whether co-transfection difficulties result from poor internalisation or the inability of DNA to escape from polyplexes. The results demonstrate the utility of this facile approach to label polyplexes without interfering with gene expression, and enable high-throughput screening of transfection reagents for achieving optimal co-transfection.

Original languageEnglish
Pages (from-to)20053-20057
Number of pages5
JournalRSC Advances
Volume9
Issue number35
DOIs
Publication statusPublished - 1 Jul 2019

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Semiconductor quantum dots
Labels
Plasmids
DNA
Throughput
Nucleic acids
Gene expression
Screening
Genes
Proteins
Imaging techniques
Nucleic Acids
Polymers

Cite this

Kretzmann, J. A., Feng, R. L., Munshi, A. M., Ho, D., Ranieri, A. M., Massi, M., ... Iyer, K. S. (2019). A facile methodology using quantum dot multiplex labels for tracking co-transfection. RSC Advances, 9(35), 20053-20057. https://doi.org/10.1039/c9ra03518d
Kretzmann, Jessica A. ; Feng, Rui Lu ; Munshi, Alaa M. ; Ho, Diwei ; Ranieri, Anna M. ; Massi, Massimiliano ; Saunders, Martin ; Norret, Marck ; Evans, Cameron W. ; Iyer, K. Swaminathan. / A facile methodology using quantum dot multiplex labels for tracking co-transfection. In: RSC Advances. 2019 ; Vol. 9, No. 35. pp. 20053-20057.
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A facile methodology using quantum dot multiplex labels for tracking co-transfection. / Kretzmann, Jessica A.; Feng, Rui Lu; Munshi, Alaa M.; Ho, Diwei; Ranieri, Anna M.; Massi, Massimiliano; Saunders, Martin; Norret, Marck; Evans, Cameron W.; Iyer, K. Swaminathan.

In: RSC Advances, Vol. 9, No. 35, 01.07.2019, p. 20053-20057.

Research output: Contribution to journalArticle

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Kretzmann JA, Feng RL, Munshi AM, Ho D, Ranieri AM, Massi M et al. A facile methodology using quantum dot multiplex labels for tracking co-transfection. RSC Advances. 2019 Jul 1;9(35):20053-20057. https://doi.org/10.1039/c9ra03518d