title: Charge-altering releasable transporters (CARTs) for the delivery and release of mRNA in living animals.
abstract: Functional delivery of mRNA to tissues in the body is key to implementing fundamentally new and potentially transformative strategies for vaccination, protein replacement therapy, and genome editing, collectively affecting approaches for the prevention, detection, and treatment of disease. Broadly applicable tools for the efficient delivery of mRNA into cultured cells would advance many areas of research, and effective and safe in vivo mRNA delivery could fundamentally transform clinical practice. Here we report the step-economical synthesis and evaluation of a tunable and effective class of synthetic biodegradable materials: charge-altering releasable transporters (CARTs) for mRNA delivery into cells. CARTs are structurally unique and operate through an unprecedented mechanism, serving initially as oligo(α-amino ester) cations that complex, protect, and deliver mRNA and then change physical properties through a degradative, charge-neutralizing intramolecular rearrangement, leading to intracellular release of functional mRNA and highly efficient protein translation. With demonstrated utility in both cultured cells and animals, this mRNA delivery technology should be broadly applicable to numerous research and therapeutic applications.
authors.affiliation: Department of Chemistry, Stanford University, Stanford, CA 94305.
authors.forename: Colin J
authors.initials: CJ
authors.lastname: McKinlay
authors.affiliation: Department of Chemistry, Stanford University, Stanford, CA 94305.
authors.forename: Jessica R
authors.initials: JR
authors.lastname: Vargas
authors.affiliation: Department of Chemistry, Stanford University, Stanford, CA 94305.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Blake
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, CA 94305.|Molecular Imaging Program at Stanford, Stanford University, Stanford, CA 94305.
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Hardy
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, CA 94305.|Molecular Imaging Program at Stanford, Stanford University, Stanford, CA 94305.
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Kanada
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, CA 94305; ccontag@stanford.edu wenderp@stanford.edu waymouth@stanford.edu.|Molecular Imaging Program at Stanford, Stanford University, Stanford, CA 94305.|Department of Microbiology & Immunology, Stanford University, Stanford, CA 94305.|Department of Bioengineering, Stanford University, Stanford, CA 94305.|Department of Radiology, Stanford University, Stanford, CA 94305.
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Contag
authors.affiliation: Department of Chemistry, Stanford University, Stanford, CA 94305; ccontag@stanford.edu wenderp@stanford.edu waymouth@stanford.edu.|Department of Chemical and Systems Biology, Stanford University, Stanford, CA 94305.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Wender
authors.affiliation: Department of Chemistry, Stanford University, Stanford, CA 94305; ccontag@stanford.edu wenderp@stanford.edu waymouth@stanford.edu.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Waymouth
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000008:administration & dosage*; D002232:Carbocyanines; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D005260:Female; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000008:administration & dosage*
chemical_list: D001672:Biocompatible Materials; D002232:Carbocyanines; D012333:RNA, Messenger; C085321:cyanine dye 5; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: cell-penetrating; gene therapy; nanoparticle; organocatalysis; stimuli-responsive