Journal article
The infectious BAC genomic DNA expression library: a high capacity vector system for functional genomics
- Abstract:
- Gene dosage plays a critical role in a range of cellular phenotypes, yet most cellular expression systems use heterologous cDNA-based vectors which express proteins well above physiological levels. In contrast, genomic DNA expression vectors generate physiologically-relevant levels of gene expression by carrying the whole genomic DNA locus of a gene including its regulatory elements. Here we describe the first genomic DNA expression library generated using the high-capacity herpes simplex virus-1 amplicon technology to deliver bacterial artificial chromosomes (BACs) into cells by viral transduction. The infectious BAC (iBAC) library contains 184,320 clones with an average insert size of 134.5 kb. We show in a Chinese hamster ovary (CHO) disease model cell line and mouse embryonic stem (ES) cells that this library can be used for genetic rescue studies in a range of contexts including the physiological restoration of Ldlr deficiency, and viral receptor expression. The iBAC library represents an important new genetic analysis tool openly available to the research community.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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- Publisher copy:
- 10.1038/srep28644
Authors
+ Biotechnology and Biological Sciences Research Council
More from this funder
- Funding agency for:
- Wade-Martins, R
- Grant:
- BB/D012910/1
- Publisher:
- Nature Publishing Group: Open Access Journals - Option C
- Journal:
- PLoS ONE More from this journal
- Volume:
- 6
- Article number:
- 28644
- Publication date:
- 2016-06-29
- Acceptance date:
- 2016-05-13
- DOI:
- ISSN:
-
2045-2322
- Pubs id:
-
pubs:629473
- UUID:
-
uuid:43618ae5-ae88-4fa7-8c04-59fcc7a00ae4
- Local pid:
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pubs:629473
- Source identifiers:
-
629473
- Deposit date:
-
2016-06-23
Terms of use
- Copyright holder:
- Lufino et al
- Copyright date:
- 2016
- Notes:
- © 2016 Lufino et al. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
- Licence:
- CC Attribution (CC BY)
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