Engineering Mosaic T7 Bacteriophages for Dual Peptide Display to Enable Cellular Targeting and Ligand-Directed Protein Capture.
4 affiliations
- 1Laboratory of Precision and Nanomedicine, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
- 2National Institute of Research and Innovation, Riga, Latvia.
- 3Institute of Technology, University of Tartu, Tartu, Estonia.
- 4Materials Research Laboratory, University of California, Santa Barbara, California, USA.
Abstract
Integrating molecular recognition, tracing, and interactome capture within a single nanoparticle could allow for probing, manipulation, and control of complex biological systems. Achieving such multifunctionality within a single well-defined nanostructure remains challenging. T7 bacteriophage-based biological nanoparticles provide a robust and genetically programmable scaffold; however, conventional T7 display systems typically accommodate only one exogenous peptide species per phage particle, limiting multifunctional and combinatorial applications. Here, we describe a mosaic phage platform that enables the simultaneous display of two peptides on separate T7 coat protein subunits. By combining genome-encoded and plasmid-driven expression of peptide-fused capsid proteins, the system provides a modular route for assembling multifunctional phage nanoparticles. We generated mosaic T7 phages co-presenting the neuropilin-1 (NRP-1)-binding peptide RPARPAR alongside either a HA-tag or a Twin-Strep (Tstrep) tag affinity handle. These engineered phages specifically target NRP-1-expressing cells and enable immunofluorescence visualization and pull-down of NRP-1 from complex lysates. Furthermore, a random peptide T7 phage library co-displaying the constant lung-homing peptide RPARPAR shows preferential pulmonary accumulation in vivo, demonstrating utility for combinatorial targeting and tissue-directed phage selection. Collectively, this dual-display approach expands phage display by enabling cell targeting, visualization, molecular interactome probing, and in vivo tissue homing within a single genetically encoded nanoparticle platform.
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BibTeX
@article{marchetti2026engineering,
title = {{Engineering Mosaic T7 Bacteriophages for Dual Peptide Display to Enable Cellular Targeting and Ligand-Directed Protein Capture.}},
author = {Luca Marchetti and Kristina Põšnograjeva and Prakash Lingasamy and Karlis Pleiko and Ivar Ilves and Tambet Teesalu},
journal = {Biotechnology journal},
year = {2026},
month = oct,
doi = {10.1002/biot.70322},
url = {https://europepmc.org/article/MED/42952180},
note = {CC BY},
}
RIS
TY - JOUR
TI - Engineering Mosaic T7 Bacteriophages for Dual Peptide Display to Enable Cellular Targeting and Ligand-Directed Protein Capture.
AU - Luca Marchetti
AU - Kristina Põšnograjeva
AU - Prakash Lingasamy
AU - Karlis Pleiko
AU - Ivar Ilves
AU - Tambet Teesalu
PY - 2026
DA - 2026/10/28
JO - Biotechnology journal
DO - 10.1002/biot.70322
UR - https://europepmc.org/article/MED/42952180
AB - Integrating molecular recognition, tracing, and interactome capture within a single nanoparticle could allow for probing, manipulation, and control of complex biological systems. Achieving such multifunctionality within a single well-defined nanostructure remains challenging. T7 bacteriophage-based biological nanoparticles provide a robust and genetically programmable scaffold; however, conventional T7 display systems typically accommodate only one exogenous peptide species per phage particle, limiting multifunctional and combinatorial applications. Here, we describe a mosaic phage platform that enables the simultaneous display of two peptides on separate T7 coat protein subunits. By combining genome-encoded and plasmid-driven expression of peptide-fused capsid proteins, the system provides a modular route for assembling multifunctional phage nanoparticles. We generated mosaic T7 phages co-presenting the neuropilin-1 (NRP-1)-binding peptide RPARPAR alongside either a HA-tag or a Twin-Strep (Tstrep) tag affinity handle. These engineered phages specifically target NRP-1-expressing cells and enable immunofluorescence visualization and pull-down of NRP-1 from complex lysates. Furthermore, a random peptide T7 phage library co-displaying the constant lung-homing peptide RPARPAR shows preferential pulmonary accumulation in vivo, demonstrating utility for combinatorial targeting and tissue-directed phage selection. Collectively, this dual-display approach expands phage display by enabling cell targeting, visualization, molecular interactome probing, and in vivo tissue homing within a single genetically encoded nanoparticle platform.
SN - 1860-6768
SN - 1860-7314
ER -