Large BACs transfect more efficiently in circular topology

Yin Cheng Wong; Andrew Osahor; Farooq Omar Maan Al-Ajli; Kumaran Narayanan

doi:10.1016/j.ab.2021.114324

Large BACs transfect more efficiently in circular topology

Yin Cheng Wong
, Andrew Osahor
, Farooq Omar Maan Al-Ajli
, Kumaran Narayanan

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

The effect of DNA topology on transfection efficiency of mammalian cells has been widely tested on plasmids smaller than 10 kb, but little is known for larger DNA vectors carrying intact genomic DNA containing introns, exons, and regulatory regions. Here, we demonstrate that circular BACs transfect more efficiently than covalently closed linear BACs. We found up to 3.1- and 8.9- fold higher eGFP expression from circular 11 kb and 100 kb BACs, respectively, compared to linear BACs. These findings provide insights for improved vector development for gene delivery and expression studies of large intact transgenes in mammalian cells.

Original language	English
Article number	114324
Journal	Analytical Biochemistry
Volume	630
DOIs	https://doi.org/10.1016/j.ab.2021.114324
State	Published - 1 Oct 2021
Externally published	Yes

Keywords

BACs
Intact
Mammalian
Topology
Transfection

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10.1016/j.ab.2021.114324

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title = "Large BACs transfect more efficiently in circular topology",

abstract = "The effect of DNA topology on transfection efficiency of mammalian cells has been widely tested on plasmids smaller than 10 kb, but little is known for larger DNA vectors carrying intact genomic DNA containing introns, exons, and regulatory regions. Here, we demonstrate that circular BACs transfect more efficiently than covalently closed linear BACs. We found up to 3.1- and 8.9- fold higher eGFP expression from circular 11 kb and 100 kb BACs, respectively, compared to linear BACs. These findings provide insights for improved vector development for gene delivery and expression studies of large intact transgenes in mammalian cells.",

keywords = "BACs, Intact, Mammalian, Topology, Transfection",

author = "Wong, \{Yin Cheng\} and Andrew Osahor and Al-Ajli, \{Farooq Omar Maan\} and Kumaran Narayanan",

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doi = "10.1016/j.ab.2021.114324",

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AU - Wong, Yin Cheng

AU - Osahor, Andrew

AU - Al-Ajli, Farooq Omar Maan

AU - Narayanan, Kumaran

PY - 2021/10/1

Y1 - 2021/10/1

N2 - The effect of DNA topology on transfection efficiency of mammalian cells has been widely tested on plasmids smaller than 10 kb, but little is known for larger DNA vectors carrying intact genomic DNA containing introns, exons, and regulatory regions. Here, we demonstrate that circular BACs transfect more efficiently than covalently closed linear BACs. We found up to 3.1- and 8.9- fold higher eGFP expression from circular 11 kb and 100 kb BACs, respectively, compared to linear BACs. These findings provide insights for improved vector development for gene delivery and expression studies of large intact transgenes in mammalian cells.

AB - The effect of DNA topology on transfection efficiency of mammalian cells has been widely tested on plasmids smaller than 10 kb, but little is known for larger DNA vectors carrying intact genomic DNA containing introns, exons, and regulatory regions. Here, we demonstrate that circular BACs transfect more efficiently than covalently closed linear BACs. We found up to 3.1- and 8.9- fold higher eGFP expression from circular 11 kb and 100 kb BACs, respectively, compared to linear BACs. These findings provide insights for improved vector development for gene delivery and expression studies of large intact transgenes in mammalian cells.

KW - BACs

KW - Intact

KW - Mammalian

KW - Topology

KW - Transfection

UR - https://www.scopus.com/pages/publications/85112015558

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DO - 10.1016/j.ab.2021.114324

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SN - 0003-2697

VL - 630

JO - Analytical Biochemistry

JF - Analytical Biochemistry

M1 - 114324

ER -

Large BACs transfect more efficiently in circular topology

Abstract

Keywords

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