TY - JOUR
T1 - Reversible Protection and Targeted Delivery of DNA Origami with a Disulfide-Containing Cationic Polymer
AU - Youssef, Sarah
AU - Tsang, Emily
AU - Samanta, Anirban
AU - Kumar, Vipin
AU - Gothelf, Kurt V
PY - 2024/3
Y1 - 2024/3
N2 - DNA nanostructures have considerable biomedical potential as intracellular delivery vehicles as they are highly homogeneous and can be functionalized with high spatial resolution. However, challenges like instability under physiological conditions, limited cellular uptake, and lysosomal degradation limit their use. This paper presents a bio-reducible, cationic polymer poly(cystaminebisacrylamide-1,6-diaminohexane) (PCD) as a reversible DNA origami protector. PCD displays a stronger DNA affinity than other cationic polymers. DNA nanostructures with PCD protection are shielded from low salt conditions and DNase I degradation and show a 40-fold increase in cell-association when linked to targeting antibodies. Confocal microscopy reveals a potential secondary cell uptake mechanism, directly delivering the nanostructures to the cytoplasm. Additionally, PCD can be removed by cleaving its backbone disulfides using the intracellular reductant, glutathione. Finally, the application of these constructs is demonstrated for targeted delivery of a cytotoxic agent to cancer cells, which efficiently decreases their viability. The PCD protective agent that is reported here is a simple and efficient method for the stabilization of DNA origami structures. With the ability to deprotect the DNA nanostructures upon entry of the intracellular space, the possibility for the use of DNA origami in pharmaceutical applications is enhanced.
AB - DNA nanostructures have considerable biomedical potential as intracellular delivery vehicles as they are highly homogeneous and can be functionalized with high spatial resolution. However, challenges like instability under physiological conditions, limited cellular uptake, and lysosomal degradation limit their use. This paper presents a bio-reducible, cationic polymer poly(cystaminebisacrylamide-1,6-diaminohexane) (PCD) as a reversible DNA origami protector. PCD displays a stronger DNA affinity than other cationic polymers. DNA nanostructures with PCD protection are shielded from low salt conditions and DNase I degradation and show a 40-fold increase in cell-association when linked to targeting antibodies. Confocal microscopy reveals a potential secondary cell uptake mechanism, directly delivering the nanostructures to the cytoplasm. Additionally, PCD can be removed by cleaving its backbone disulfides using the intracellular reductant, glutathione. Finally, the application of these constructs is demonstrated for targeted delivery of a cytotoxic agent to cancer cells, which efficiently decreases their viability. The PCD protective agent that is reported here is a simple and efficient method for the stabilization of DNA origami structures. With the ability to deprotect the DNA nanostructures upon entry of the intracellular space, the possibility for the use of DNA origami in pharmaceutical applications is enhanced.
KW - DNA origami
KW - antibody
KW - bioreducible polymers
KW - cationic polymers
KW - cell uptake
KW - Nanostructures/chemistry
KW - DNA/chemistry
KW - Disulfides/chemistry
KW - Cations/chemistry
KW - Nucleic Acid Conformation
KW - Polymers/chemistry
UR - http://www.scopus.com/inward/record.url?scp=85175376204&partnerID=8YFLogxK
U2 - 10.1002/smll.202301058
DO - 10.1002/smll.202301058
M3 - Journal article
C2 - 37916910
SN - 1613-6810
VL - 20
JO - Small
JF - Small
IS - 10
M1 - 2301058
ER -