TY - JOUR
T1 - Deconvoluting the Effect of Cell-Penetrating Peptides for Enhanced and Controlled Insertion of Large-Scale DNA Nanopores
AU - Zhang, Xialin
AU - Malle, Mette Galsgaard
AU - Thomsen, Rasmus P.
AU - Sørensen, Rasmus Schøler
AU - Sørensen, Emily Winther
AU - Hatzakis, Nikos S.
AU - Kjems, Jørgen
N1 - Publisher Copyright:
© 2024 The Authors. Published by American Chemical Society.
PY - 2024/4/17
Y1 - 2024/4/17
N2 - DNA nanopores have emerged as powerful tools for molecular sensing, but the efficient insertion of large DNA nanopores into lipid membranes remains challenging. In this study, we investigate the potential of cell-penetrating peptides (CPPs), specifically SynB1 and GALA, to enhance the insertion efficiency of large DNA nanopores. We constructed SynB1- or GALA-functionalized DNA nanopores with an 11 nm inner diameter and visualized and quantified their membrane insertion using a TIRF microscopy-based single-liposome assay. The results demonstrated that incorporating an increasing number of SynB1 or GALA peptides into the DNA nanopore significantly enhanced the membrane perforation. Kinetic analysis revealed that the DNA nanopore scaffold played a role in prearranging the CPPs, which facilitated membrane interaction and pore formation. Notably, the use of pH-responsive GALA peptides allowed highly efficient and pH-controlled insertion of large DNA pores. Furthermore, single-channel recording elucidated that the insertion process of single GALA-modified nanopores into planar lipid bilayers was dynamic, likely forming transient large toroidal pores. Overall, our study highlights the potential of CPPs as insertion enhancers for DNA nanopores, which opens avenues for improved molecule sensing and the controlled release of cargo molecules.
AB - DNA nanopores have emerged as powerful tools for molecular sensing, but the efficient insertion of large DNA nanopores into lipid membranes remains challenging. In this study, we investigate the potential of cell-penetrating peptides (CPPs), specifically SynB1 and GALA, to enhance the insertion efficiency of large DNA nanopores. We constructed SynB1- or GALA-functionalized DNA nanopores with an 11 nm inner diameter and visualized and quantified their membrane insertion using a TIRF microscopy-based single-liposome assay. The results demonstrated that incorporating an increasing number of SynB1 or GALA peptides into the DNA nanopore significantly enhanced the membrane perforation. Kinetic analysis revealed that the DNA nanopore scaffold played a role in prearranging the CPPs, which facilitated membrane interaction and pore formation. Notably, the use of pH-responsive GALA peptides allowed highly efficient and pH-controlled insertion of large DNA pores. Furthermore, single-channel recording elucidated that the insertion process of single GALA-modified nanopores into planar lipid bilayers was dynamic, likely forming transient large toroidal pores. Overall, our study highlights the potential of CPPs as insertion enhancers for DNA nanopores, which opens avenues for improved molecule sensing and the controlled release of cargo molecules.
KW - bioconjugation
KW - cell-penetrating peptides
KW - DNA nanopore
KW - insertion enhancement
KW - single-liposome assay
KW - TIRF
KW - DNA/chemistry
KW - Lipid Bilayers/chemistry
KW - Cell-Penetrating Peptides
KW - Kinetics
KW - Nanopores
UR - http://www.scopus.com/inward/record.url?scp=85189993215&partnerID=8YFLogxK
U2 - 10.1021/acsami.3c18636
DO - 10.1021/acsami.3c18636
M3 - Journal article
C2 - 38573069
AN - SCOPUS:85189993215
SN - 1944-8244
VL - 16
SP - 18422
EP - 18433
JO - A C S Applied Materials and Interfaces
JF - A C S Applied Materials and Interfaces
IS - 15
ER -