Mechanisms and optimization of in vivo delivery of lipophilic siRNAs
Abstract
Cholesterol-conjugated siRNAs can silence gene expression in vivo. Here we synthesize a variety of lipophilic siRNAs and use them to elucidate the requirements for siRNA delivery in vivo. We show that conjugation to bile acids and long-chain fatty acids, in addition to cholesterol, mediates siRNA uptake into cells and gene silencing in vivo. Efficient and selective uptake of these siRNA conjugates depends on interactions with lipoprotein particles, lipoprotein receptors and transmembrane proteins. High-density lipoprotein (HDL) directs siRNA delivery into liver, gut, kidney and steroidogenic organs, whereas low-density lipoprotein (LDL) targets siRNA primarily to the liver. LDL-receptor expression is essential for siRNA delivery by LDL particles, and SR-BI receptor expression is required for uptake of HDL-bound siRNAs. Cellular uptake also requires the mammalian homolog of the Caenorhabditis elegans transmembrane protein Sid1. Our results demonstrate that conjugation to lipophilic molecules enables effective siRNA uptake through a common mechanism that can be exploited to optimize therapeutic siRNA delivery.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
L2-siRNA shows greater cell internalization than unconjugated parent siRNA in neuroblastoma cells, while Chol-siRNA shows the highest uptake in serum-free media.
“There was more cell internalization of L2-siRNA than the parent siRNA, while Chol-siRNA, a benchmark chosen for its known cell penetration capacity [10], had the highest level of uptake in serum-free media (Supplementary Fig. S1A).”
From Elucidating brain transport pathways and cell type-dependent gene silencing of a durable lipid–siRNA conjugate administered into cerebrospinal fluid (2025) · cites this paper as [10]
Conjugation of siRNA to lipophilic moieties is intended to improve cellular uptake through membrane interactions.
“Conjugation of siRNA to lipophilic moieties is intended to improve cellular uptake through membrane interactions [9, 10].”
From Elucidating brain transport pathways and cell type-dependent gene silencing of a durable lipid–siRNA conjugate administered into cerebrospinal fluid (2025) · cites this paper as [10]
Chol-siRNA is a benchmark chosen for its known cell penetration capacity.
“while Chol-siRNA, a benchmark chosen for its known cell penetration capacity [10], had the highest level of uptake in serum-free media”
From Elucidating brain transport pathways and cell type-dependent gene silencing of a durable lipid–siRNA conjugate administered into cerebrospinal fluid (2025) · cites this paper as [10]
Cites
A Receptor-Mediated Pathway for Cholesterol Homeostasis
Brown, M. S., & Goldstein, J. L. (1986) · Science · doi:10.1126/science.3513311 · Cited by 5,703 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Non-viral vectors for gene-based therapy
Yin, H., Kanasty, R. L., Eltoukhy, A. A., Vegas, A. J., Dorkin, J. R., & Anderson, D. G. (2014) · Nature Reviews Genetics · doi:10.1038/nrg3763 · Cited by 3,123 · see citing works
Knocking down barriers: advances in siRNA delivery
Whitehead, K. A., Langer, R., & Anderson, D. G. (2009) · Nature Reviews Drug Discovery · doi:10.1038/nrd2742 · Cited by 3,115 · see citing works
MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins
Vickers, K. C., Palmisano, B. T., Shoucri, B. M., Shamburek, R. D., & Remaley, A. T. (2011) · Nature Cell Biology · doi:10.1038/ncb2210 · Cited by 2,741 · see citing works
In vivo genome editing using Staphylococcus aureus Cas9
Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche, B., Shalem, O., Wu, X., Makarova, K. S., et al. (2015) · Nature · doi:10.1038/nature14299 · Cited by 2,687 · see citing works
Delivery materials for siRNA therapeutics
Kanasty, R., Dorkin, J. R., Vegas, A., & Anderson, D. (2013) · Nature Materials · doi:10.1038/nmat3765 · Cited by 1,800 · see citing works
Gilleron, J., Querbes, W., Zeigerer, A., Borodovsky, A., Marsico, G., Schubert, U., Manygoats, K., Seifert, S., Andree, C., Stöter, M., et al. (2013) · Nature Biotechnology · doi:10.1038/nbt.2612 · Cited by 1,681 · see citing works
Self-citationNair, J. K., Willoughby, J. L. S., Chan, A., Charisse, K., Alam, M. R., Wang, Q., Hoekstra, M., Kandasamy, P., Kel’in, A. V., Milstein, S., et al. (2014) · Journal of the American Chemical Society · doi:10.1021/ja505986a · Cited by 1,141 · see citing works
Self-citationDeveloping mRNA-vaccine technologies
Schlake, T., Thess, A., Fotin-Mleczek, M., & Kallen, K.-J. (2012) · RNA Biology · doi:10.4161/rna.22269 · Cited by 622 · see citing works
A holistic approach to targeting disease with polymeric nanoparticles
Cheng, C. J., Tietjen, G. T., Saucier-Sawyer, J. K., & Saltzman, W. M. (2015) · Nature Reviews Drug Discovery · doi:10.1038/nrd4503 · Cited by 480 · see citing works
Expanding RNAi therapeutics to extrahepatic tissues with lipophilic conjugates
Brown, K. M., Nair, J. K., Janas, M. M., Anglero-Rodriguez, Y. I., Dang, L. T. H., Peng, H., Theile, C. S., Castellanos-Rizaldos, E., Brown, C., Foster, D., et al. (2022) · Nature Biotechnology · doi:10.1038/s41587-022-01334-x · Cited by 258 · see citing works
Self-citationNonviral methods for siRNA delivery
Gao, K., & Huang, L. (2009) · Molecular Pharmaceutics · doi:10.1021/mp800134q · Cited by 244 · see citing works
Wang, L. L., Liu, Y., Chung, J. J., Wang, T., Gaffey, A. C., Lu, M., Cavanaugh, C. A., Zhou, S., Kanade, R., Atluri, P., et al. (2017) · Nature Biomedical Engineering · doi:10.1038/s41551-017-0157-y · Cited by 239 · see citing works
Lipid nanoparticles for short interfering RNA delivery
Leung, A. K. K., Tam, Y. Y. C., & Cullis, P. R. (2014) · Advances in genetics · doi:10.1016/b978-0-12-800148-6.00004-3 · Cited by 158 · see citing works
Therapeutic efficacy in a hemophilia B model using a biosynthetic mRNA liver depot system
DeRosa, F., Guild, B., Karve, S., Smith, L., Love, K., Dorkin, J. R., Kauffman, K. J., Zhang, J., Yahalom, B., Anderson, D. G., et al. (2016) · Gene Therapy · doi:10.1038/gt.2016.46 · Cited by 159 · see citing works
Improving siRNA Delivery <i>In Vivo</i> Through Lipid Conjugation
Osborn, M. F., & Khvorova, A. (2018) · Nucleic Acid Therapeutics · doi:10.1089/nat.2018.0725 · Cited by 137 · see citing works
Visualization of self-delivering hydrophobically modified siRNA cellular internalization
Ly, S., Navaroli, D. M., Didiot, M.-C., Cardia, J., Pandarinathan, L., Alterman, J. F., Fogarty, K., Standley, C., Lifshitz, L. M., Bellve, K. D., et al. (2017) · Nucleic Acids Research · doi:10.1093/nar/gkw1005 · Cited by 132 · see citing works
Nikan, M., Osborn, M. F., Coles, A. H., Godinho, B. M., Hall, L. M., Haraszti, R. A., Hassler, M. R., Echeverria, D., Aronin, N., & Khvorova, A. (2016) · Molecular Therapy - Nucleic Acids · doi:10.1038/mtna.2016.50 · Cited by 85 · see citing works
RNA delivery biomaterials for the treatment of genetic and rare diseases
Zhao, W., Hou, X., Vick, O. G., & Dong, Y. (2019) · Biomaterials · doi:10.1016/j.biomaterials.2019.119291 · Cited by 82 · see citing works
A Mirror Image Fluorogenic Aptamer Sensor for Live-Cell Imaging of MicroRNAs
Zhong, W., & Sczepanski, J. T. (2019) · ACS Sensors · doi:10.1021/acssensors.9b00252 · Cited by 55 · see citing works
Bortolozzi, A., Manashirov, S., Chen, A., & Artigas, F. (2021) · Pharmacology & Therapeutics · doi:10.1016/j.pharmthera.2021.107873 · Cited by 37 · see citing works
Polymeric micelles for siRNA delivery
Falamarzian, A., Xiong, X.-B., Uludag, H., & Lavasanifar, A. (2012) · Journal of Drug Delivery Science and Technology · doi:10.1016/s1773-2247(12)50004-3 · Cited by 25 · see citing works
Sorets, A. G., Schwensen, K. R., Francini, N., Kjar, A., Abdulrahman, A. M., Shostak, A., Katdare, K. A., Schoch, K. M., Cowell, R. P., Park, J. C., et al. (2025) · Nucleic Acids Research · doi:10.1093/nar/gkaf600 · Cited by 6 · see citing works
“Conjugation of siRNA to lipophilic moieties is intended to improve cellular uptake through membrane interactions [ 9 , 10 ].”
Sorets, A., Schwensen, K., Francini, N., Kjar, A., Abdulrahman, A., Shostak, A., Katdare, K., Schoch, K., Cowell, R., Park, J., et al. (2024) · doi:10.1101/2024.06.09.598079 · Cited by 4 · see citing works
- Authors
- Wolfrum, C., Shi, S., Jayaprakash, K. N., Jayaraman, M., Wang, G., Pandey, R. K., Rajeev, K. G., Nakayama, T., Charrise, K., Ndungo, E. M., Zimmermann, T., Koteliansky, V., Manoharan, M., & Stoffel, M.
- Affiliations
- ETH Zurich (CH)
- Rockefeller University (US)
- Alnylam Pharmaceuticals (United States) (US)
- Corresponding author
- Stoffel, Markus
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2007-09-16
- Volume
- 25
- Issue
- 10
- Pages
- 1149-1157
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W2156955518
- Cited by
- 974
- References
- 1
- Citations by year
- 2026: 28 · 2025: 44 · 2024: 40 · 2023: 51 · 2022: 46 · 2021: 45 · 2020: 55 · 2019: 53 · 2018: 37 · 2017: 38 · 2016: 57 · 2015: 57 · 2014: 57 · 2013: 64 · 2012: 70
- FWCI
- 20.12
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of General Medical Sciences (P01GM073047)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Keywords
- siRNA delivery, cholesterol conjugation, bile acid conjugation, lipoprotein receptors, high-density lipoprotein, low-density lipoprotein, LDL receptor, siRNA uptake, liver targeting
- MeSH terms
- Animals, Apolipoproteins B, Cholesterol, Cricetinae, Lipoproteins, HDL, Lipoproteins, LDL, Mesocricetus, Gene Transfer Techniques, Receptors, Lipoprotein, RNA Interference, RNA, Small Interfering, Mice
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
