β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides
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 enzym...
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MDPI AG
2018-07-01
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| Series: | Biomolecules |
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| Online Access: | http://www.mdpi.com/2218-273X/8/3/51 |
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doaj-ca42dfa2a83b4d0c880a789631e2dd6c2020-11-24T23:07:59ZengMDPI AGBiomolecules2218-273X2018-07-01835110.3390/biom8030051biom8030051β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating PeptidesShane R. Stone0Tatjana Heinrich1Suzy M. Juraja2Jiulia N. Satiaputra3Clinton M. Hall4Mark Anastasas5Anna D. Mills6Christopher A. Chamberlain7Scott Winslow8Kristin Priebatsch9Paula T. Cunningham10Katrin Hoffmann11Nadia Milech12Telethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaTelethon Kids Institute, University of Western Australia, Subiaco, WA 6008, AustraliaThe 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.http://www.mdpi.com/2218-273X/8/3/51cell penetrating peptidehigh-throughput screeningfunctional validationcell internalizationcytosolic delivery |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Shane R. Stone Tatjana Heinrich Suzy M. Juraja Jiulia N. Satiaputra Clinton M. Hall Mark Anastasas Anna D. Mills Christopher A. Chamberlain Scott Winslow Kristin Priebatsch Paula T. Cunningham Katrin Hoffmann Nadia Milech |
| spellingShingle |
Shane R. Stone Tatjana Heinrich Suzy M. Juraja Jiulia N. Satiaputra Clinton M. Hall Mark Anastasas Anna D. Mills Christopher A. Chamberlain Scott Winslow Kristin Priebatsch Paula T. Cunningham Katrin Hoffmann Nadia Milech β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides Biomolecules cell penetrating peptide high-throughput screening functional validation cell internalization cytosolic delivery |
| author_facet |
Shane R. Stone Tatjana Heinrich Suzy M. Juraja Jiulia N. Satiaputra Clinton M. Hall Mark Anastasas Anna D. Mills Christopher A. Chamberlain Scott Winslow Kristin Priebatsch Paula T. Cunningham Katrin Hoffmann Nadia Milech |
| author_sort |
Shane R. Stone |
| title |
β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides |
| title_short |
β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides |
| title_full |
β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides |
| title_fullStr |
β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides |
| title_full_unstemmed |
β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides |
| title_sort |
β-lactamase tools for establishing cell internalization and cytosolic delivery of cell penetrating peptides |
| publisher |
MDPI AG |
| series |
Biomolecules |
| issn |
2218-273X |
| publishDate |
2018-07-01 |
| description |
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. |
| topic |
cell penetrating peptide high-throughput screening functional validation cell internalization cytosolic delivery |
| url |
http://www.mdpi.com/2218-273X/8/3/51 |
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