alexa An Alternate Method for Efficient Delivery of Catalyzing Enzymes
ISSN: 2168-9547

Molecular Biology: Open Access
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Research Article

An Alternate Method for Efficient Delivery of Catalyzing Enzymes

Uma Lakshmipathy1* Andrew Fontes1, Michael Poderycki2 and Jonathan D. Chesnut2
1Cell Biology & Stem Cell Sciences, Life Technologies, 5781 Van Allen Way, Carlsbad, CA 92008, USA
2Synthetic Biology Systems, Life Technologies, 5781 Van Allen Way, Carlsbad, CA 92008, USA
Corresponding Author : Uma Lakshmipathy
Cell Biology & Stem Cell Sciences Life Technologies
5781 Van Allen Way, Carlsbad, CA 92008, USA
E-mail: [email protected]
Received September 12, 2013; Accepted October 19, 2013; Published October 26, 2013
Citation: Lakshmipathy U, Fontes A, Poderycki M, Chesnut JD (2013) An Alternate Method for Efficient Delivery of Catalyzing Enzymes. Mol Biol 2:112. doi:10.4172/2168-9547.1000112
Copyright: © 2013 Lakshmipathy U, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Improved cellular engineering tools have enabled scientists to modify genomic sequences at a new pace for a variety of applications. However, in order to create more accurate tools for site specific targeting and editing, improved delivery systems for modifying enzymes are required. These enzymes allow end users to disrupt, insert or edit genes utilizing sequence specific homology. Current applications require co-transfection of an engineering plasmid in tandem with the vector carrying the modifying enzyme. However, co-transfection can negatively affect transfection efficiencies as well as increase probability of randomly integrating DNA. Here we report the transient delivery of catalyzing enzymes via BacMam non-integrating virus at high efficiency. Baculovirus is a non-infectious delivery system that can transduce a variety of mammalian cells. We have shown improved delivery of NLS-Cre recombinase for the excision of regions in disrupted genes to enable expression. In addition, we have tested PhiC31 integrase delivery for the guided insertion of plasmid DNA to a specific locus. The transient expression provided by BacMam, coupled with ease of transfection, creates an ideal system for delivery of catalyzing enzymes in mammalian cells.

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