Combinatorial approach to determine functional group effects on lipidoid-mediated siRNA delivery.
The application of RNA interference (RNAi), either in the clinic or in the laboratory, requires safe and effective delivery methods. Here, we develop a combinatorial approach to synthesize a library of delivery vectors based on two lipid-like substrates with known siRNA delivery capabilities. Members of this library have a mixture of lipid-like tails and feature appendages containing hydroxyl, carbamate, ether, or amine functional groups as well as variations in alkyl chain length and branching. Using a luciferase reporter system in HeLa cells, we studied the relationship between lipid chemical modification and delivery performance in vitro. The impact of the functional group was shown to vary depending on the overall amine content and tail number of the delivery vector. Additionally, in vivo performance was evaluated using a Factor VII knockdown assay. Two library members, each containing ether groups, were found to knock down the target protein at levels comparable to those of the parent delivery vector. These results demonstrate that small chemical changes to the delivery vector impact knockdown efficiency and cell viability both in vitro and in vivo. The work described here identifies new materials for siRNA delivery and provides new insight into the parameters for optimized chemical makeup of lipid-like siRNA delivery materials.
Mahon
Kerry P
KP
Department of Chemical Engineering and David H. Koch Institute of Integrative Cancer Research, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
Love
Kevin T
KT
Whitehead
Kathryn A
KA
Qin
June
J
Akinc
Akin
A
Leshchiner
Elizaveta
E
Leshchiner
Ignaty
I
Langer
Robert
R
Anderson
Daniel G
DG
D000178:Acrylamides / Q000737:chemistry; D000179:Acrylates / Q000737:chemistry; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D020650:Combinatorial Chemistry Techniques; D004305:Dose-Response Relationship, Drug; D005167:Factor VII / Q000032:analysis; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000138:chemical synthesis / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D015394:Molecular Structure; D034741:RNA, Small Interfering / Q000737:chemistry / Q000494:pharmacology*
D000178:Acrylamides; D000179:Acrylates; D008055:Lipids; D034741:RNA, Small Interfering; D005167:Factor VII