A universal molecular control for DNA, mRNA and protein expression

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 compat...

Celý popis

Uloženo v:
Podrobná bibliografie
Vydáno v:Nature communications Ročník 15; číslo 1; s. 2480 - 13
Hlavní autoři: Gunter, Helen M., Youlten, Scott E., Reis, Andre L. M., McCubbin, Tim, Madala, Bindu Swapna, Wong, Ted, Stevanovski, Igor, Cipponi, Arcadi, Deveson, Ira W., Santini, Nadia S., Kummerfeld, Sarah, Croucher, Peter I., Marcellin, Esteban, Mercer, Tim R.
Médium: Journal Article
Jazyk:angličtina
Vydáno: London Nature Publishing Group UK 20.03.2024
Nature Publishing Group
Nature Portfolio
Témata:
38/23
38/91
42/40
631/1647/48
631/1647/514/2254
631/337/475
631/61/185
631/61/212/2019
82/58
82/75
82/80
Biology
Deoxyribonucleic acid
DNA
DNA - genetics
DNA sequencing
Gene expression
Genes
Genomics
Humanities and Social Sciences
Mass spectrometry
Mass spectroscopy
Molecular biology
multidisciplinary
Next-generation sequencing
Proteins
Proteomics
Ribonucleic acid
RNA
RNA, Messenger - genetics
RNA, Messenger - metabolism
Science
Science (multidisciplinary)
Scientific imaging
Transcriptomics
Translation
ISSN:2041-1723, 2041-1723
On-line přístup:Získat plný text
Tagy: Přidat tag
Žádné tagy, Buďte první, kdo vytvoří štítek k tomuto záznamu!
Popis
Shrnutí: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. Multi-omics analyses powerfully combine gene expression and translation, however no available controls can be used across these techniques. Here the authors develop pREF, a universal control construct designed for use in DNA, RNA and protein analyses.
Bibliografie:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 14
content type line 23
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-024-46456-9