Absolute measurement of gene transcripts with Selfie-digital PCR

Abstract Absolute measurement of the number of RNA transcripts per gene is necessary to compare gene transcription among different tissues or experimental conditions and to assess transcription of genes that have a variable copy number per cell such as mitochondrial DNA. Here, we present a method ca...

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Autores principales:	Petar Podlesniy, Ramon Trullas
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/44c1b2302d1d46bc9a6983bbdbf07262
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spelling	oai:doaj.org-article:44c1b2302d1d46bc9a6983bbdbf072622021-12-02T12:32:07ZAbsolute measurement of gene transcripts with Selfie-digital PCR10.1038/s41598-017-08270-w2045-2322https://doaj.org/article/44c1b2302d1d46bc9a6983bbdbf072622017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08270-whttps://doaj.org/toc/2045-2322Abstract Absolute measurement of the number of RNA transcripts per gene is necessary to compare gene transcription among different tissues or experimental conditions and to assess transcription of genes that have a variable copy number per cell such as mitochondrial DNA. Here, we present a method called Selfie-digital PCR that measures the absolute amount of an RNA transcript produced by its own coding DNA at a particular moment. Overcoming the limitations of previous approaches, Selfie-digital PCR allows for the quantification of nuclear and mitochondrial gene transcription in a strand-specific manner that is comparable among tissues and cell types that differ in gene copy number or metabolic state. Using Selfie-digital PCR, we found that, with the exception of the liver, different organs exhibit marked variations in mitochondrial DNA copy number but similar transcription of mitochondrial DNA heavy and light chains, thus suggesting a preferential role of mitochondrial DNA abundance over its transcription in organ function. Moreover, the strand-specific analysis of mitochondrial transcription afforded by Selfie-digital PCR showed that transcription of the heavy strand was significantly higher than that of the light strand in all the tissues studied.Petar PodlesniyRamon TrullasNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-15 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Petar Podlesniy Ramon Trullas Absolute measurement of gene transcripts with Selfie-digital PCR
description	Abstract Absolute measurement of the number of RNA transcripts per gene is necessary to compare gene transcription among different tissues or experimental conditions and to assess transcription of genes that have a variable copy number per cell such as mitochondrial DNA. Here, we present a method called Selfie-digital PCR that measures the absolute amount of an RNA transcript produced by its own coding DNA at a particular moment. Overcoming the limitations of previous approaches, Selfie-digital PCR allows for the quantification of nuclear and mitochondrial gene transcription in a strand-specific manner that is comparable among tissues and cell types that differ in gene copy number or metabolic state. Using Selfie-digital PCR, we found that, with the exception of the liver, different organs exhibit marked variations in mitochondrial DNA copy number but similar transcription of mitochondrial DNA heavy and light chains, thus suggesting a preferential role of mitochondrial DNA abundance over its transcription in organ function. Moreover, the strand-specific analysis of mitochondrial transcription afforded by Selfie-digital PCR showed that transcription of the heavy strand was significantly higher than that of the light strand in all the tissues studied.
format	article
author	Petar Podlesniy Ramon Trullas
author_facet	Petar Podlesniy Ramon Trullas
author_sort	Petar Podlesniy
title	Absolute measurement of gene transcripts with Selfie-digital PCR
title_short	Absolute measurement of gene transcripts with Selfie-digital PCR
title_full	Absolute measurement of gene transcripts with Selfie-digital PCR
title_fullStr	Absolute measurement of gene transcripts with Selfie-digital PCR
title_full_unstemmed	Absolute measurement of gene transcripts with Selfie-digital PCR
title_sort	absolute measurement of gene transcripts with selfie-digital pcr
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/44c1b2302d1d46bc9a6983bbdbf07262
work_keys_str_mv	AT petarpodlesniy absolutemeasurementofgenetranscriptswithselfiedigitalpcr AT ramontrullas absolutemeasurementofgenetranscriptswithselfiedigitalpcr
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Absolute measurement of gene transcripts with Selfie-digital PCR

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