β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides
AbstractThe 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
- Supplementary File 1:
PDF-Document (PDF, 1634 KB)
Share & Cite This Article
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.
Stone SR, Heinrich T, Juraja SM, Satiaputra JN, Hall CM, Anastasas M, Mills AD, Chamberlain CA, Winslow S, Priebatsch K, Cunningham PT, Hoffmann K, Milech N. β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides. Biomolecules. 2018; 8(3):51.Chicago/Turabian Style
Stone, Shane R.; Heinrich, Tatjana; Juraja, Suzy M.; Satiaputra, Jiulia N.; Hall, Clinton M.; Anastasas, Mark; Mills, Anna D.; Chamberlain, Christopher A.; Winslow, Scott; Priebatsch, Kristin; Cunningham, Paula T.; Hoffmann, Katrin; Milech, Nadia. 2018. "β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides." Biomolecules 8, no. 3: 51.
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.