CoChat
Sign Up
Cited by 123RNA Interference and Gene DeliveryRetinal Development and DisordersAdvanced Drug Delivery Systems

Lipid nanoparticles with PEG-variant surface modifications mediate genome editing in the mouse retina

Open access
Gautam, Milan, Jozic, Antony, Su, Grace Li-Na, Herrera-Barrera, Marco, Curtis, Allison, Arrizabalaga, Sebastian, Tschetter, Wayne, Ryals, Renee C. +1 more·2023·Nature Communications
DOI 10.1038/s41467-023-42189-3PMC PMC10575971
Open this paper in CoChat — free accountOpen PDF

Abstract

Ocular delivery of lipid nanoparticle (LNPs) packaged mRNA can enable efficient gene delivery and editing. We generated LNP variants through the inclusion of positively charged-amine-modified polyethylene glycol (PEG)-lipids (LNPa), negatively charged-carboxyl-(LNPz) and carboxy-ester (LNPx) modified PEG-lipids, and neutral unmodified PEG-lipids (LNP). Subretinal injections of LNPa containing Cre mRNA in the mouse show tdTomato signal in the retinal pigmented epithelium (RPE) like conventional LNPs. Unexpectedly, LNPx and LNPz show 27% and 16% photoreceptor transfection, respectively, with striking localization extending from the photoreceptor synaptic pedicle to the outer segments, displaying pan-retinal distribution in the photoreceptors and RPE. LNPx containing Cas9 mRNA and sgAi9 leads to the formation of an oval elongated structure with a neutral charge resulting in 16.4% editing restricted to RPE. Surface modifications of LNPs with PEG variants can alter cellular tropism of mRNA. LNPs enable genome editing in the retina and in the future can be used to correct genetic mutations that lead to blindness.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/s41467-023-42189-3

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

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?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

The citation lists haven’t been fetched yet

References 41 · Cited by 123

Research works printed but not linked: 35

  1. [1]Lee, E. K. in Inherited Retinal Disease (ed. Yu, H.-G.) 21–38 (Springer Nature, 2022). 10.1007/978-981-16-7337-5_2. · DOI
  2. [2]Mustafi D Hisama FM Huey J Chao JR The current state of genetic testing platforms for inherited retinal diseases Ophthalmol. Retin. 2022 6 702 710 10.1016/j.oret.2022.03.011 PMC9356993 35307606 · DOI
  3. [3]Darrow JJ Luxturna: FDA documents reveal the value of a costly gene therapy Drug Discov. Today 2019 24 949 954 10.1016/j.drudis.2019.01.019 30711576 · DOI
  4. [4]Chirco KR Martinez C Lamba DA Advancements in pre-clinical development of gene editing-based therapies to treat inherited retinal diseases Vis. Res. 2023 209 108257 10.1016/j.visres.2023.108257 37210864 PMC10524382 · DOI
  5. [5]Fenner BJ Gene-based therapeutics for inherited retinal diseases Front. Genet. 2022 12 794805 10.3389/fgene.2021.794805 35069693 PMC8782148 · DOI
  6. [6]Gerasimavicius L Livesey BJ Marsh JA Loss-of-function, gain-of-function and dominant-negative mutations have profoundly different effects on protein structure Nat. Commun. 2022 13 3895 10.1038/s41467-022-31686-6 35794153 PMC9259657 · DOI
  7. [7]Xi Z Vats A Sahel J-A Chen Y Byrne LC Gene augmentation prevents retinal degeneration in a CRISPR/Cas9-based mouse model of PRPF31 retinitis pigmentosa Nat. Commun. 2022 13 7695 10.1038/s41467-022-35361-8 36509783 PMC9744804 · DOI
  8. [8]Han, I. C. et al. Characterization of a novel Pde6b-deficient rat model of retinal degeneration and treatment with adeno-associated virus (AAV) gene therapy. Gene Ther. 1–7 (2022) 10.1038/s41434-022-00365-y. 36175490 · DOI
  9. [9]Yin J Safeguarding genome integrity during gene-editing therapy in a mouse model of age-related macular degeneration Nat. Commun. 2022 13 7867 10.1038/s41467-022-35640-4 36550137 PMC9780342 · DOI
  10. [10]Maeder ML Development of a gene-editing approach to restore vision loss in Leber congenital amaurosis type 10 Nat. Med. 2019 25 229 233 10.1038/s41591-018-0327-9 30664785 · DOI · Paper page
  11. [11]Pfromm JK Bonillo M Dauletbekov D Bucher K Fischer MD Plasmid-mediated gene transfer of Cas9 induces vector-related but not SpCas9-related immune responses in human retinal pigment epithelial cells Sci. Rep. 2022 12 13202 10.1038/s41598-022-17269-x 35915300 PMC9343442 · DOI
  12. [12]Hanlon KS High levels of AAV vector integration into CRISPR-induced DNA breaks Nat. Commun. 2019 10 4439 10.1038/s41467-019-12449-2 31570731 PMC6769011 · DOI · Paper page
  13. [14]Kazemian P Lipid-nanoparticle-based delivery of CRISPR/Cas9 genome-editing components Mol. Pharm. 2022 19 1669 1686 10.1021/acs.molpharmaceut.1c00916 35594500 PMC9176214 · DOI
  14. [15]Barbier AJ Jiang AY Zhang P Wooster R Anderson DG The clinical progress of mRNA vaccines and immunotherapies Nat. Biotechnol. 2022 40 840 854 10.1038/s41587-022-01294-2 35534554 · DOI · Paper page
  15. [16]Prakash G Microfluidic fabrication of lipid nanoparticles for the delivery of nucleic acids Adv. Drug Deliv. Rev. 2022 184 114197 10.1016/j.addr.2022.114197 35288219 PMC9035142 · DOI
  16. [18]Kim J Rapid generation of circulating and mucosal decoy human ACE2 using mRNA nanotherapeutics for the potential treatment of SARS-CoV-2 Adv. Sci. 2022 9 2202556 10.1002/advs.202202556 PMC9762296 36216580 · DOI
  17. [20]Mastrocola, F., Nunes, F. & Sgobbi, P. in Amyloidosis and Fabry Disease: A Clinical Guide (eds. Xavier de Ávila, D. & Villacorta Junior, H.) 235–276 (Springer International Publishing, 2023). 10.1007/978-3-031-17759-0_22. · DOI
  18. [21]Patel S Ryals RC Weller KK Pennesi ME Sahay G Lipid nanoparticles for delivery of messenger RNA to the back of the eye J. Control. Release 2019 303 91 100 10.1016/j.jconrel.2019.04.015 30986436 PMC6579630 · DOI · Paper page
  19. [22]Herrera-Barrera M Peptide-guided lipid nanoparticles deliver mRNA to the neural retina of rodents and nonhuman primates Sci. Adv. 2023 9 eadd4623 10.1126/sciadv.add4623 36630502 PMC9833661 · DOI
  20. [23]Eygeris Y Patel S Jozic A Sahay G Deconvoluting lipid nanoparticle structure for messenger RNA delivery Nano Lett. 2020 20 4543 4549 10.1021/acs.nanolett.0c01386 32375002 PMC7228479 · DOI
  21. [24]Nguyen Le,N-M Charge-converting nanoemulsions as promising retinal drug and gene delivery systems ACS Appl. Mater. Interfaces 2022 14 44981 44991 10.1021/acsami.2c11649 36125912 PMC9542710 · DOI
  22. [25]Herrera M Kim J Eygeris Y Jozic A Sahay G Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery Biomater. Sci. 2021 9 4289 4300 10.1039/D0BM01947J 33586742 PMC8769212 · DOI
  23. [26]Madisen L A robust and high-throughput Cre reporting and characterization system for the whole mouse brain Nat. Neurosci. 2010 13 133 140 10.1038/nn.2467 20023653 PMC2840225 · DOI · Paper page
  24. [27]Makino CL Recoverin regulates light-dependent phosphodiesterase activity in retinal rods J. Gen. Physiol. 2004 123 729 741 10.1085/jgp.200308994 15173221 PMC2234569 · DOI
  25. [29]Wei T Cheng Q Min Y-L Olson EN Siegwart DJ Systemic nanoparticle delivery of CRISPR-Cas9 ribonucleoproteins for effective tissue specific genome editing Nat. Commun. 2020 11 3232 10.1038/s41467-020-17029-3 32591530 PMC7320157 · DOI
  26. [30]Qiu M Lipid nanoparticle-mediated codelivery of Cas9 mRNA and single-guide RNA achieves liver-specific in vivo genome editing of Angptl3 Proc. Natl Acad. Sci. 2021 118 e2020401118 10.1073/pnas.2020401118 33649229 PMC7958351 · DOI
  27. [31]Samaridou E Heyes J Lutwyche P Lipid nanoparticles for nucleic acid delivery: current perspectives Adv. Drug Deliv. Rev. 2020 154–155 37 63 10.1016/j.addr.2020.06.002 32526452 · DOI
  28. [32]Anand P Stahel VP The safety of Covid-19 mRNA vaccines: a review Patient Saf. Surg. 2021 15 20 10.1186/s13037-021-00291-9 33933145 PMC8087878 · DOI
  29. [33]Pickens CJ Johnson SN Pressnall MM Leon MA Berkland CJ Practical considerations, challenges, and limitations of bioconjugation via azide–alkyne cycloaddition Bioconjug. Chem. 2018 29 686 701 10.1021/acs.bioconjchem.7b00633 29287474 PMC6310217 · DOI
  30. [35]Guimaraes PPG Ionizable lipid nanoparticles encapsulating barcoded mRNA for accelerated in vivo delivery screening J. Control. Release 2019 316 404 417 10.1016/j.jconrel.2019.10.028 31678653 PMC7032071 · DOI
  31. [37]Sun D Non-viral gene therapy for stargardt disease with ECO/pRHO-ABCA4 self-assembled nanoparticles Mol. Ther. 2020 28 293 303 10.1016/j.ymthe.2019.09.010 31611143 PMC6952167 · DOI
  32. [38]Shen J Suprachoroidal gene transfer with nonviral nanoparticles Sci. Adv. 2020 6 eaba1606 10.1126/sciadv.aba1606 32937452 PMC7458446 · DOI · Paper page
  33. [39]Palumbo MC MicroRNA137-loaded lipid nanoparticles regulate synaptic proteins in the prefrontal cortex Mol. Ther. 2023 S1525-0016 00448 3 PMC10556225 37644723
  34. [40]Herrera-Barrera M Lipid nanoparticle-enabled intracellular delivery of prime editors AAPS J. 2023 25 65 10.1208/s12248-023-00833-2 37380935 PMC13198972 · DOI
  35. [41]Boatright JH Methodologies for analysis of patterning in the mouse RPE sheet Mol. Vis. 2015 21 40 60 25593512 PMC4301600

Authors
Gautam, M., Jozic, A., Su, G. L.-N., Herrera-Barrera, M., Curtis, A., Arrizabalaga, S., Tschetter, W., Ryals, R. C., & Sahay, G.
Affiliations
  • Oregon State University (US)
  • Oregon Health & Science University (US)
Corresponding author
  • Ryals, Renee C.
  • Sahay, Gaurav
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2023-10-13
Volume
14
Issue
1
Pages
6468-6468
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-023-42189-3
PMC
PMC10575971
OpenAlex
W4387609025
Cited by
123
References
41
Citations by year
2026: 31 · 2025: 60 · 2024: 29 · 2023: 3
FWCI
10.96
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
U.S. Department of Health & Human Services | NIH | National Eye Institute (1R01EY033423-01A1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advanced Drug Delivery Systems
Keywords
lipid nanoparticles, ocular gene delivery, genome editing, mRNA delivery, retinal photoreceptors, retinal pigment epithelium, subretinal injection, Cas9, single guide RNA, cell tropism, blindness
MeSH terms
Gene Editing, Animals, Lipids, Liposomes, Polyethylene Glycols, RNA, Messenger, RNA, Small Interfering, Mice, Nanoparticles, Retinal Pigment Epithelium

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