Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs

Bo Yu; Xinmei Wang; Chenguang Zhou; Lesheng Teng; Wei Ren; Zhaogang Yang; Chih Hsin Shih; Tianyou Wang; Robert J. Lee; Suoqin Tang; L. James Lee

doi:10.1007/s11095-014-1366-7

Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs

Bo Yu, Xinmei Wang, Chenguang Zhou, Lesheng Teng, Wei Ren, Zhaogang Yang, Chih Hsin Shih, Tianyou Wang, Robert J. Lee, Suoqin Tang, L. James Lee

Research output: Contribution to journal › Article › peer-review

56 Scopus citations

Abstract

Purpose: Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and microRNA (miRNA).

Method: In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.

Results: We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.

Conclusions: Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.

Original language	English (US)
Pages (from-to)	2685-2695
Number of pages	11
Journal	Pharmaceutical Research
Volume	31
Issue number	10
DOIs	https://doi.org/10.1007/s11095-014-1366-7
State	Published - Apr 17 2014
Externally published	Yes

Keywords

intracellular trafficking
lipid nanoparticles
miRNA
molecular beacon
siRNA

ASJC Scopus subject areas

Biotechnology
Molecular Medicine
Pharmacology
Pharmaceutical Science
Organic Chemistry
Pharmacology (medical)

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10.1007/s11095-014-1366-7

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title = "Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs",

abstract = "Purpose: Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and microRNA (miRNA).Method: In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.Results: We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.Conclusions: Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.",

keywords = "intracellular trafficking, lipid nanoparticles, miRNA, molecular beacon, siRNA",

author = "Bo Yu and Xinmei Wang and Chenguang Zhou and Lesheng Teng and Wei Ren and Zhaogang Yang and Shih, {Chih Hsin} and Tianyou Wang and Lee, {Robert J.} and Suoqin Tang and Lee, {L. James}",

note = "Funding Information: This work was supported by NSF Nanoscale Science and Engineering Center (NSEC) grant EEC-0914790 and the Chinese National 863 Project (No. 2012AA020804). Publisher Copyright: {\textcopyright} 2014 Springer Science+Business Media New York.",

year = "2014",

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doi = "10.1007/s11095-014-1366-7",

language = "English (US)",

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T1 - Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs

AU - Yu, Bo

AU - Wang, Xinmei

AU - Zhou, Chenguang

AU - Teng, Lesheng

AU - Ren, Wei

AU - Yang, Zhaogang

AU - Shih, Chih Hsin

AU - Wang, Tianyou

AU - Lee, Robert J.

AU - Tang, Suoqin

AU - Lee, L. James

N1 - Funding Information: This work was supported by NSF Nanoscale Science and Engineering Center (NSEC) grant EEC-0914790 and the Chinese National 863 Project (No. 2012AA020804). Publisher Copyright: © 2014 Springer Science+Business Media New York.

PY - 2014/4/17

Y1 - 2014/4/17

N2 - Purpose: Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and microRNA (miRNA).Method: In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.Results: We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.Conclusions: Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.

AB - Purpose: Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and microRNA (miRNA).Method: In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.Results: We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.Conclusions: Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.

KW - intracellular trafficking

KW - lipid nanoparticles

KW - miRNA

KW - molecular beacon

KW - siRNA

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Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs

Abstract

Keywords

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