TY - JOUR
T1 - Tracking the DNA complexation state of pBAE polyplexes in cells with super resolution microscopy
AU - Riera, Roger
AU - Feiner-Gracia, Natalia
AU - Fornaguera, Cristina
AU - Cascante, Anna
AU - Borrós, Salvador
AU - Albertazzi, Lorenzo
N1 - Funding Information:
The authors thanks the Spanish Ministry of Economy, Industry and Competitiveness, through Project SAF2016-75241-R, as part of the Generalitat de Catalunya through the CERCA program. L. A. thanks the European Research Council (ERC-StG-757397).
Publisher Copyright:
© 2019 The Royal Society of Chemistry.
PY - 2019/10/14
Y1 - 2019/10/14
N2 - The future of gene therapy relies on the development of efficient and safe delivery vectors. Poly(β-amino ester)s are promising cationic polymers capable of condensing oligonucleotides into nanoparticles-polyplexes-and deliver them into the cell nucleus, where the gene material would be expressed. The complexation state during the crossing of biological barriers is crucial: Polymers should tightly complex DNA before internalization and then release to allow free DNA to reach the nucleus. However, measuring the complexation state in cells is challenging due to the nanometric size of polyplexes and the difficulties to study the two components (polymer and DNA) independently. Here we propose a method to visualize and quantify the two components of a polyplex inside cells, with nanometre scale resolution, using two-colour direct stochastic reconstruction super-resolution microscopy (dSTORM). With our approach, we tracked the complexation state of pBAE polyplexes from cell binding to DNA release and nuclear entry revealing time evolution and the final fate of DNA and pBAE polymers in mammalian cells.
AB - The future of gene therapy relies on the development of efficient and safe delivery vectors. Poly(β-amino ester)s are promising cationic polymers capable of condensing oligonucleotides into nanoparticles-polyplexes-and deliver them into the cell nucleus, where the gene material would be expressed. The complexation state during the crossing of biological barriers is crucial: Polymers should tightly complex DNA before internalization and then release to allow free DNA to reach the nucleus. However, measuring the complexation state in cells is challenging due to the nanometric size of polyplexes and the difficulties to study the two components (polymer and DNA) independently. Here we propose a method to visualize and quantify the two components of a polyplex inside cells, with nanometre scale resolution, using two-colour direct stochastic reconstruction super-resolution microscopy (dSTORM). With our approach, we tracked the complexation state of pBAE polyplexes from cell binding to DNA release and nuclear entry revealing time evolution and the final fate of DNA and pBAE polymers in mammalian cells.
UR - http://www.scopus.com/inward/record.url?scp=85072945113&partnerID=8YFLogxK
U2 - 10.1039/c9nr02858g
DO - 10.1039/c9nr02858g
M3 - Article
C2 - 31552987
AN - SCOPUS:85072945113
SN - 2040-3364
VL - 11
SP - 17869
EP - 17877
JO - Nanoscale
JF - Nanoscale
IS - 38
ER -