Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapieDec 4, 2025

Peptides help deliver large CRISPR gene-editing tools without viruses

mRNA Technology Weekly Brief ↗PubMed ↗DOI ↗

Updated Jan 7, 2026

Abstract

The peptide-assisted lipofection (PAL) system achieved 98.7% transfection efficiency in HEK293T cells for large genetic constructs.

PAL successfully delivered plasmids up to 29 kb while maintaining high cell viability.
Indel formation efficiency of 44.1% was observed in gene editing experiments using PAL.
Fluorescence microscopy confirmed PAL's effective endosomal escape and nuclear targeting.
In vivo studies in mouse models indicated sustained gene expression in liver tissue with PAL.
PAL outperformed conventional plasmid delivery methods, suggesting it may provide a safer alternative to viral vectors.

Simplified

Efficient and safe delivery of large genetic constructs such as CRISPR/Cas9 plasmids remains a critical bottleneck in gene therapy. In this work, the peptide-assisted lipofection (PAL) system was developed as a breakthrough non-viral vector for gene delivery, specifically designed for large plasmids including CRISPR/Cas9 constructs. This system achieved exceptional transfection efficiency up to 98.7 % in HEK293T cells, surpassing conventional delivery methods such as electroporation and standard lipofections. PAL successfully delivered large plasmids up to 29 kb while maintaining high cell viability and achieved 44.1 % indel formation efficiency in gene editing experiments. Fluorescence microscopy verified PAL's efficient endosomal escape and nuclear targeting abilities. The superior performance of the PAL is attributed to its cellular uptake and endosomal escape enhanced by transfection-assisting peptide. In vivo studies in mouse models showed sustained gene expression in liver tissue, demonstrating superior performance compared to naked plasmid delivery. These results establish PAL as a versatile and promising platform for gene therapy and genome editing, offering a safer alternative to viral vectors for large genetic payload delivery.

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Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Byeong Hee Hwang has patent pending to Incheon National University. Yeong Chae Ryu has patent pending to Incheon National University. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

PubMed

Peptides help deliver large CRISPR gene-editing tools without viruses

Abstract

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Funding

Competing interests

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