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Review
. 2025 Oct;43(10):2399-2409.
doi: 10.1016/j.tibtech.2025.05.002. Epub 2025 May 28.

Scaling DNA engineering

Weiyi Li  1 , Po-Hsiang Hung  2 , Takeshi Matsui  2 , Sasha F Levy  2 , Gavin Sherlock  3
Affiliations
Review

Scaling DNA engineering

Weiyi Li et al. Trends Biotechnol. 2025 Oct.

Abstract

DNA can be engineered to produce new biologics, gene therapies, and cellular therapies, and to reprogram organisms. Having the ability to engineer DNA at scale can accelerate the development of these applications. Existing technologies excel at short oligonucleotide synthesis by chemical or enzymatic methods (up to 2000 bp) and intermediate-size DNA assembly (up to 5-7 kb). Yet synthesizing sequence-validated longer DNA (>10 kb) and/or constructing highly complex combinatorial DNA libraries at scale remains a significant challenge, due largely to technical and cost barriers. Inspired by recent studies on an in vivo DNA processing platform for megabase-long DNA assembly and on high-throughput sequence verification, we discuss how these platforms may be used to achieve DNA engineering at scale.

Keywords: DNA assembly; bacterial conjugation; homologous recombination.

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Conflict of interest statement

Declaration of interests W.L. and S.F.L. are co-authors on patent applications based on the methods presented in this work, specifically BPS and SCRIVENER. S.F.L and G.S. co-founded BacStitch DNA Inc., to commercialize BPS and SCRIVENER. P.H.H., T.M., G.S., and S.F.L have financial interest in BacStitch DNA, Inc.

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