Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/142406
Title: Tracking genome-editing and associated molecular perturbations by SWATH mass spectrometry
Authors: Lin, Qifeng
Low, Larry W. L.
Lau, Adam
Chua, Esther W. L.
Matsuoka, Yuji
Lian, Yilong
Monteiro, Antónia
Tate, Stephen
Gunaratne, Jayantha
Carney, Tom J.
Keywords: Science::Medicine
Issue Date: 2019
Source: Lin, Q., Low, L. W. L., Lau, A., Chua, E. W. L., Matsuoka, Y., Lian, Y., . . . Carney, T. J. (2019). Tracking genome-editing and associated molecular perturbations by SWATH mass spectrometry. Scientific Reports, 9(1), 15240-. doi:10.1038/s41598-019-51612-z
Journal: Scientific Reports
Abstract: Advances in gene editing now allow reverse genetics to be applied to a broad range of biological systems. Ultimately, any modification to coding sequences requires confirmation at the protein level, although immunoblotting is often hampered by antibody quality or availability especially in non-model species. Sequential Window Acquisition of All Theoretical Spectra (SWATH), a mass spectrometry (MS) technology with exceptional quantitative reproducibility and accuracy, offers an ideal alternative for protein-based confirmation. Here, using genome edits in mouse, zebrafish and Bicyclus anynana butterflies produced using either homologous recombination or targeted nucleases, we demonstrate absence of the targeted proteins using SWATH, thus confirming successful editing. We show that SWATH is a robust antibody-independent alternative for monitoring gene editing at the protein level and broadly applicable across diverse organisms and targeted genome manipulation techniques. Moreover, SWATH concomitantly defines the global proteome response in the edited organism, which may provide pertinent biological insights.
URI: https://hdl.handle.net/10356/142406
ISSN: 2045-2322
DOI: 10.1038/s41598-019-51612-z
Rights: © 2019 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Te images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:LKCMedicine Journal Articles

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