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Biomolecules 2018, 8(3), 51; https://doi.org/10.3390/biom8030051

β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides

1
Telethon Kids Institute, University of Western Australia, Subiaco, WA 6008, Australia
2
Phylogica Pty Ltd., Subiaco, WA 6008, Australia
*
Author to whom correspondence should be addressed.
Received: 11 May 2018 / Revised: 22 June 2018 / Accepted: 29 June 2018 / Published: 11 July 2018
(This article belongs to the Special Issue Cell Penetrating Peptides)
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Abstract

The ability of cell penetrating peptides (CPPs) to deliver biologically relevant cargos into cells is becoming more important as targets in the intracellular space continue to be explored. We have developed two assays based on CPP-dependent, intracellular delivery of TEM-1 β-lactamase enzyme, a functional biological molecule comparable in size to many protein therapeutics. The first assay focuses on the delivery of full-length β-lactamase to evaluate the internalization potential of a CPP sequence. The second assay uses a split-protein system where one component of β-lactamase is constitutively expressed in the cytoplasm of a stable cell line and the other component is delivered by a CPP. The delivery of a split β-lactamase component evaluates the cytosolic delivery capacity of a CPP. We demonstrate that these assays are rapid, flexible and have potential for use with any cell type and CPP sequence. Both assays are validated using canonical and novel CPPs, with limits of detection from <500 nM to 1 µM. Together, the β-lactamase assays provide compatible tools for functional characterization of CPP activity and the delivery of biological cargos into cells. View Full-Text
Keywords: cell penetrating peptide; high-throughput screening; functional validation; cell internalization; cytosolic delivery cell penetrating peptide; high-throughput screening; functional validation; cell internalization; cytosolic delivery
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Stone, S.R.; Heinrich, T.; Juraja, S.M.; Satiaputra, J.N.; Hall, C.M.; Anastasas, M.; Mills, A.D.; Chamberlain, C.A.; Winslow, S.; Priebatsch, K.; Cunningham, P.T.; Hoffmann, K.; Milech, N. β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides. Biomolecules 2018, 8, 51.

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