Enhanced mRNA delivery into lymphocytes enabled by lipid-varied libraries of charge-altering releasable transporters
Year: 2018
Type: article
Abstract: We report a strategy for generating a combinatorial library of oligonucleotide transporters with varied lipid domains and their use in the efficient transfection of lymphocytes with mRNA in vitro and in vivo. This library is based on amphiphilic charge-altering releasable transporters (CARTs) that contain a lipophilic block functionalized with various side-chain lipids and a polycationic α-amino ester mRNA-binding block that undergoes rearrangement to neutral small molecules, resulting in mRNA release. We show that certain binary mixtures of these lipid-varied CARTs provide up to a ninefold enhancement in mRNA translation in lymphocytes in vitro relative to either a single-lipid CART component alone or the commercial reagent Lipofectamine 2000, corresponding to a striking increase in percent transfection from 9-12% to 80%. Informed by the results with binary mixtures, we further show that CARTs consisting of optimized ratios of the two lead lipids incorporated into a single hybrid-lipid transporter molecule maintain the same delivery efficacy as the noncovalent mixture of two CARTs. The lead lipid CART mixtures and hybrid-lipid CARTs show enhanced lymphocyte transfection in primary T cells and in vivo in mice. This combinatorial approach for rapidly screening mRNA delivery vectors has provided lipid-varied CART mixtures and hybrid-lipid CARTs that exhibit significant improvement in mRNA delivery to lymphocytes, a finding of potentially broad value in research and clinical applications.
Institutions: Stanford University, Stanford Medicine, Stanford Cancer Institute
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: SYNTHETIC STUDIES ON TUMOR PROMOTERS AND INHIBITORS, Synthetic Studies Related to Cancer Research/Treatment, GOALI: SusChem: Organocatalysis: A Platform for Sustainable Polymer Chemistry, BD Canto II RUO Cytometer, SELECTIVE CATALYTIC OXIDATIONS: OPPORTUNITIES AND CHALLENGES FOR SELECTIVE CONVERSION OF RENEWABLE RESOURCE, Synthetic Studies on Tumor Promoters and Inhibitors, G9000245758, G9000247180, Synthetic Studies Related to Cancer Research/Treatment
OpenAlex ID: W2808372952