A universal molecular control for DNA, mRNA and protein expression.

Autor:	Gunter HM; Australian Institute of Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, Australia.; BASE mRNA Facility, The University of Queensland, Brisbane, Queensland, Australia.; ARC Centre of Excellence in Synthetic Biology, The University of Queensland, Brisbane, Queensland, Australia., Youlten SE; Department of Genetics, Yale University School of Medicine, New Haven, CT, 06510, USA.; Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; St Vincent's Clinical School, University of New South Wales, Sydney, New South Wales, Australia., Reis ALM; Genomics and Inherited Disease Program, Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; Centre for Population Genomics, Garvan Institute of Medical Research and Murdoch Children's Research Institute, Sydney, New South Wales, Australia.; School of Electrical and Information Engineering, University of Sydney, Sydney, New South Wales, Australia., McCubbin T; Australian Institute of Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, Australia.; ARC Centre of Excellence in Synthetic Biology, The University of Queensland, Brisbane, Queensland, Australia., Madala BS; Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; Centre for Population Genomics, Garvan Institute of Medical Research and Murdoch Children's Research Institute, Sydney, New South Wales, Australia., Wong T; Garvan Institute of Medical Research, Sydney, New South Wales, Australia., Stevanovski I; Genomics and Inherited Disease Program, Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; Centre for Population Genomics, Garvan Institute of Medical Research and Murdoch Children's Research Institute, Sydney, New South Wales, Australia., Cipponi A; Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; St Vincent's Clinical School, University of New South Wales, Sydney, New South Wales, Australia., Deveson IW; Genomics and Inherited Disease Program, Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; Centre for Population Genomics, Garvan Institute of Medical Research and Murdoch Children's Research Institute, Sydney, New South Wales, Australia.; School of Electrical and Information Engineering, University of Sydney, Sydney, New South Wales, Australia., Santini NS; Centro Nacional de Investigación Disciplinaria en Conservación y Mejoramiento de Ecosistemas Forestales, INIFAP, Ciudad de México, 04010, Mexico., Kummerfeld S; Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; St Vincent's Clinical School, University of New South Wales, Sydney, New South Wales, Australia., Croucher PI; Garvan Institute of Medical Research, Sydney, New South Wales, Australia.; St Vincent's Clinical School, University of New South Wales, Sydney, New South Wales, Australia., Marcellin E; Australian Institute of Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, Australia.; ARC Centre of Excellence in Synthetic Biology, The University of Queensland, Brisbane, Queensland, Australia., Mercer TR; Australian Institute of Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, Australia. t.mercer@uq.edu.au.; BASE mRNA Facility, The University of Queensland, Brisbane, Queensland, Australia. t.mercer@uq.edu.au.; ARC Centre of Excellence in Synthetic Biology, The University of Queensland, Brisbane, Queensland, Australia. t.mercer@uq.edu.au.; Garvan Institute of Medical Research, Sydney, New South Wales, Australia. t.mercer@uq.edu.au.
Jazyk:	angličtina
Zdroj:	Nature communications [Nat Commun] 2024 Mar 20; Vol. 15 (1), pp. 2480. Date of Electronic Publication: 2024 Mar 20.
DOI:	10.1038/s41467-024-46456-9
Abstrakt:	The expression of genes encompasses their transcription into mRNA followed by translation into protein. In recent years, next-generation sequencing and mass spectrometry methods have profiled DNA, RNA and protein abundance in cells. However, there are currently no reference standards that are compatible across these genomic, transcriptomic and proteomic methods, and provide an integrated measure of gene expression. Here, we use synthetic biology principles to engineer a multi-omics control, termed pREF, that can act as a universal molecular standard for next-generation sequencing and mass spectrometry methods. The pREF sequence encodes 21 synthetic genes that can be in vitro transcribed into spike-in mRNA controls, and in vitro translated to generate matched protein controls. The synthetic genes provide qualitative controls that can measure sensitivity and quantitative accuracy of DNA, RNA and peptide detection. We demonstrate the use of pREF in metagenome DNA sequencing and RNA sequencing experiments and evaluate the quantification of proteins using mass spectrometry. Unlike previous spike-in controls, pREF can be independently propagated and the synthetic mRNA and protein controls can be sustainably prepared by recipient laboratories using common molecular biology techniques. Together, this provides a universal synthetic standard able to integrate genomic, transcriptomic and proteomic methods. (© 2024. The Author(s).)
Databáze:	MEDLINE
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