Self-assembling protein nanoparticles for cytosolic delivery of nucleic acids and proteins

Feyisayo Eweje; Vanessa Ibrahim; Aram Shajii; Michelle L Walsh; Kiran Ahmad; Assma Alrefai; Dominie Miyasato; Jessie R Davis; Hyunok Ham; Kaicheng Li; Michael Roehrl; Carolyn A Haller; David R Liu; Jiaxuan Chen; Elliot L Chaikof

doi:10.1038/s41587-025-02664-2

Self-assembling protein nanoparticles for cytosolic delivery of nucleic acids and proteins

Nat Biotechnol. 2026 Apr;44(4):602-615. doi: 10.1038/s41587-025-02664-2. Epub 2025 May 15.

Authors

Feyisayo Eweje^#^{1

2

3

4}, Vanessa Ibrahim¹, Aram Shajii¹, Michelle L Walsh^{1

2

3}, Kiran Ahmad¹, Assma Alrefai¹, Dominie Miyasato¹, Jessie R Davis^{5

6

7}, Hyunok Ham^{1

4}, Kaicheng Li^{1

4}, Michael Roehrl⁸, Carolyn A Haller^{1

4}, David R Liu^{5

6

7}, Jiaxuan Chen^#^{9

10}, Elliot L Chaikof^{11

12}

Affiliations

¹ Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
² Harvard and MIT Division of Health Science and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA.
³ Harvard/MIT MD-PhD Program, Boston, MA, USA.
⁴ Wyss Institute of Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
⁵ Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
⁶ Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.
⁷ Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁸ Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
⁹ Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. jchen13@bidmc.harvard.edu.
¹⁰ Wyss Institute of Biologically Inspired Engineering, Harvard University, Boston, MA, USA. jchen13@bidmc.harvard.edu.
¹¹ Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. echaikof@bidmc.harvard.edu.
¹² Wyss Institute of Biologically Inspired Engineering, Harvard University, Boston, MA, USA. echaikof@bidmc.harvard.edu.

^# Contributed equally.

PMID: 40374955
DOI: 10.1038/s41587-025-02664-2

Abstract

Intracellular delivery of biomacromolecules is hampered by low efficiency and cytotoxicity. Here we report the development of elastin-based nanoparticles for therapeutic delivery (ENTER), a recombinant elastin-like polypeptide (ELP)-based delivery system for effective cytosolic delivery of biomacromolecules in vitro and in vivo. Through iterative design, we developed fourth-generation ELPs fused to cationic endosomal escape peptides (EEPs) that self-assemble into pH-responsive micellar nanoparticles and enable cytosolic entry of cargo following endocytic uptake. In silico screening of α-helical peptide libraries led to the discovery of an EEP (EEP13) with 48% improved protein delivery efficiency versus a benchmark peptide. Our lead ELP-EEP13 showed similar or superior performance compared to lipid-based transfection reagents in the delivery of mRNA-encoded, DNA-encoded and protein-form Cre recombinase and CRISPR gene editors as well as short interfering RNAs to multiple cell lines and primary cell types. Intranasal administration of ELP-EEP13 combined with Cre protein achieved efficient editing of lung epithelial cells in reporter mice.

MeSH terms

Animals
Cytosol* / metabolism
Drug Delivery Systems*
Elastin / chemistry
Humans
Mice
Nanoparticles* / chemistry
Nucleic Acids* / administration & dosage
Peptides / chemistry
Proteins*

Substances

Nucleic Acids
Elastin
Proteins
Peptides

Abstract

MeSH terms

Substances

Grants and funding