Dendritic peptide-conjugated polymeric nanovectors for non-toxic delivery of plasmid DNA and enhanced non-viral transfection of immune cells
Sijia Yi,
Sun-Young Kim,
Michael P. Vincent,
Simseok A. Yuk,
Sharan Bobbala,
Fanfan Du,
Evan Alexander Scott
Affiliations
Sijia Yi
Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA
Sun-Young Kim
Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA; SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University (SKKU), 2066 Seobu-ro, Jangan-gu, Suwon-si, Gyeonggi-do 16419, Republic of Korea
Michael P. Vincent
Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA
Simseok A. Yuk
Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA
Sharan Bobbala
Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA; SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University (SKKU), 2066 Seobu-ro, Jangan-gu, Suwon-si, Gyeonggi-do 16419, Republic of Korea; Department of Pharmaceutical Sciences, School of Pharmacy, West Virginia University, Morgantown, WV 26505, USA; Simpson Querrey Institute, Northwestern University, Chicago, IL 60611, USA; Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, IL 60611, USA; Department of Microbiology-Immunology, Northwestern University, Chicago, IL 60611, USA
Fanfan Du
Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA
Evan Alexander Scott
Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA; Simpson Querrey Institute, Northwestern University, Chicago, IL 60611, USA; Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, IL 60611, USA; Department of Microbiology-Immunology, Northwestern University, Chicago, IL 60611, USA; Corresponding author
Summary: Plasmid DNA (pDNA) transfection is advantageous for gene therapies requiring larger genetic elements, including “all-in-one” CRISPR/Cas9 plasmids, but is limited by toxicity as well as poor intracellular release and transfection efficiency in immune cell populations. Here, we developed a synthetic non-viral gene delivery platform composed of poly(ethylene glycol)-b-poly(propylene sulfide) copolymers linked to a cationic dendritic peptide (DP) via a reduceable bond, PEG-b-PPS-ss-DP (PPDP). A library of self-assembling PPDP polymers was synthesized and screened to identify optimal constructs capable of transfecting macrophages with small (pCMV-DsRed, 4.6 kb) and large (pL-CRISPR.EFS.tRFP, 11.7 kb) plasmids. The optimized PPDP construct transfected macrophages, fibroblasts, dendritic cells, and T cells more efficiently and with less toxicity than a commercial Lipo2K reagent, regardless of pDNA size and under standard culture conditions in the presence of serum. The PPDP technology described herein is a stimuli-responsive polymeric nanovector that can be leveraged to meet diverse challenges in gene delivery.
