Nucleic acid-mediated intracellular protein delivery by lipid-like nanoparticles.
Intracellular protein delivery has potential biotechnological and therapeutic application, but remains technically challenging. In contrast, a plethora of nucleic acid carriers have been developed, with lipid-based nanoparticles (LNPs) among the most clinically advanced reagents for oligonucleotide delivery. Here, we validate the hypothesis that oligonucleotides can serve as packaging materials to facilitate protein entrapment within and intracellular delivery by LNPs. Using two distinct model proteins, horseradish peroxidase and NeutrAvidin, we demonstrate that LNPs can yield efficient intracellular protein delivery in vitro when one or more oligonucleotides have been conjugated to the protein cargo. Moreover, in experiments with NeutrAvidin in vivo, we show that oligonucleotide conjugation significantly enhances LNP-mediated protein uptake within various spleen cell populations, suggesting that this approach may be particularly suitable for improved delivery of protein-based vaccines to antigen-presenting cells.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.
Ahmed A
AA
Eltoukhy
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.
Delai
D
Chen
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.
Omid
O
Veiseh
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.
Jeisa M
JM
Pelet
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.
Hao
H
Yin
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.
Yizhou
Y
Dong
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, United States. Electronic address: dgander@mit.edu.
Daniel G
DG
Anderson
D000818:Animals; D001360:Avidin / Q000378:metabolism*; D055592:Biophysical Phenomena; D002232:Carbocyanines / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D016503:Drug Delivery Systems; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006367:HeLa Cells; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D042541:Intracellular Space / Q000378:metabolism*; D008055:Lipids / Q000737:chemistry*; D008810:Mice, Inbred C57BL; D053758:Nanoparticles / Q000737:chemistry*; D009696:Nucleic Acids / Q000378:metabolism*; D009841:Oligonucleotides / Q000378:metabolism; D019265:Spectroscopy, Near-Infrared; D013154:Spleen / Q000378:metabolism
D002232:Carbocyanines; D008055:Lipids; D009696:Nucleic Acids; D009841:Oligonucleotides; C085321:cyanine dye 5; C093483:neutravidin; D001360:Avidin; D004247:DNA; D006735:Horseradish Peroxidase
Intracellular delivery; Lipid; Nanoparticles; Nucleic acid; Oligonucleotide; Protein delivery