Proteomics

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Tracking genome-editing and associated molecular perturbations by SWATH mass spectrometry

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-mammalian species. Here, we show that Sequential Window Acquisition of All Theoretical Spectra (SWATH) - Mass Spectrometry (MS) is a robust antibody-independent alternative for monitoring gene editing at the protein level and concomitantly defines proteome responses which may provide pertinent biological insights.

INSTRUMENT(S): TripleTOF 6600

ORGANISM(S): Bicyclus Anynana Danio Rerio (zebrafish) (brachydanio Rerio) Mus Musculus (mouse)

TISSUE(S): Pupa, Embryo, Tail Fin, Cell Culture

SUBMITTER: Qifeng Lin  

LAB HEAD: Jayantha Gunaratne

PROVIDER: PXD013176 | Pride | 2019-10-24

REPOSITORIES: Pride

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160331_IS448_Pool_IDA_2ug_1.wiff Wiff
160331_IS448_Pool_IDA_2ug_1.wiff.scan Wiff
160331_IS448_Pool_IDA_2ug_2.wiff Wiff
160331_IS448_Pool_IDA_2ug_2.wiff.scan Wiff
160331_IS448_Pool_IDA_2ug_3.wiff Wiff
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Publications

Tracking genome-editing and associated molecular perturbations by SWATH mass spectrometry.

Lin Qifeng Q   Low Larry W L LWL   Lau Adam A   Chua Esther W L EWL   Matsuoka Yuji Y   Lian Yilong Y   Monteiro Antónia A   Tate Stephen S   Gunaratne Jayantha J   Carney Tom J TJ  

Scientific reports 20191023 1

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  ...[more]

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