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Title: Expanding the DNA editing toolbox: novel lambda integrase variants targeting microalgal and human genome sequences
Authors: Siau, Jia Wei
Siddiqui, Asim Azhar
Lau, Sze Yi
Kannan, Srinivasaraghavan
Peter, Sabrina
Zeng, Yingying
Verma, Chandra
Droge, Peter
Ghadessy, John F.
Keywords: Medicine, Health and Life Sciences
Issue Date: 2024
Source: Siau, J. W., Siddiqui, A. A., Lau, S. Y., Kannan, S., Peter, S., Zeng, Y., Verma, C., Droge, P. & Ghadessy, J. F. (2024). Expanding the DNA editing toolbox: novel lambda integrase variants targeting microalgal and human genome sequences. PloS One, 19(2), e0292479-.
Project: NRF-CRP21-2018-0002 
Journal: PloS one 
Abstract: Recombinase enzymes are extremely efficient at integrating very large DNA fragments into target genomes. However, intrinsic sequence specificities curtail their use to DNA sequences with sufficient homology to endogenous target motifs. Extensive engineering is therefore required to broaden applicability and robustness. Here, we describe the directed evolution of novel lambda integrase variants capable of editing exogenous target sequences identified in the diatom Phaeodactylum tricornutum and the algae Nannochloropsis oceanica. These microorganisms hold great promise as conduits for green biomanufacturing and carbon sequestration. The evolved enzyme variants show >1000-fold switch in specificity towards the non-natural target sites when assayed in vitro. A single-copy target motif in the human genome with homology to the Nannochloropsis oceanica site can also be efficiently targeted using an engineered integrase, both in vitro and in human cells. The developed integrase variants represent useful additions to the DNA editing toolbox, with particular application for targeted genomic insertion of large DNA cargos.
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0292479
Schools: School of Biological Sciences 
Organisations: LambdaGen Pte Ltd, Singapore 
Rights: © 2024 Siau et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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