Work

Programmable large-cargo integration: Overcoming size constraints for next-generation gene therapy

Year: 2025

Type: article

Abstract: The emergence of base and prime editors-genome editing tools that avoid double-strand breaks (DSBs)-has enabled precise point mutations, insertions, inversions, deletions, and substitutions, which accelerates the development of single-intervention therapies and advances individualized genomic medicine. However, their limited efficiency in inserting large DNA fragments has restricted applications for correcting diverse pathogenic mutations within a single gene. In this review, we explore three recently developed strategies for efficient large DNA cargo insertion (>1 kb): CRISPR-associated Tn7-like transposases (CASTs), PE-integrase systems, and R2 retrotransposon fusions (nCas9-R2). We examine the applications of these systems in both bacterial and mammalian contexts and discuss their respective advantages and current limitations. Finally, we address persistent challenges and propose potential directions to guide future research.

Source: Synthetic and Systems Biotechnology

DOI: 10.1016/j.synbio.2025.11.008

Authors: Lifang Yu, Mario Andrea Marchisio

Institutions: Huangshan University, Northeastern University

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,500

Funder: Huangshan University