Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury

There is a lack of systematic research on the expression of internal control genes used for gene expression normalization in real-time reverse transcription polymerase chain reaction in spinal cord injury research. In this study, we used rat models of spinal cord hemisection to analyze the expressio...

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Autores principales:	Wei Liu, Jie Yu, Yi-Fan Wang, Qian-Qian Shan, Ya-Xian Wang
Formato:	article
Lenguaje:	EN
Publicado:	Wolters Kluwer Medknow Publications 2022
Materias:	genorm analysis; reference genes; internal control genes; normalization; normfinder analysis; reverse transcription-quantitative polymerase chain reaction; spinal cord injury; stability of gene expression Neurology. Diseases of the nervous system RC346-429
Acceso en línea:	https://doaj.org/article/2a51440108954b5f86995299df2515a5
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spelling	oai:doaj.org-article:2a51440108954b5f86995299df2515a52021-11-19T12:16:45ZSelection of suitable internal controls for gene expression normalization in rats with spinal cord injury1673-537410.4103/1673-5374.327350https://doaj.org/article/2a51440108954b5f86995299df2515a52022-01-01T00:00:00Zhttp://www.nrronline.org/article.asp?issn=1673-5374;year=2022;volume=17;issue=6;spage=1387;epage=1392;aulast=Liuhttps://doaj.org/toc/1673-5374There is a lack of systematic research on the expression of internal control genes used for gene expression normalization in real-time reverse transcription polymerase chain reaction in spinal cord injury research. In this study, we used rat models of spinal cord hemisection to analyze the expression stability of 13 commonly applied reference genes: Actb, Ankrd27, CypA, Gapdh, Hprt1, Mrpl10, Pgk1, Rictor, Rn18s, Tbp, Ubc, Ubxn11, and Ywhaz. Our results show that the expression of Ankrd27, Ubc, and Tbp were stable after spinal cord injury, while Actb was the most unstable internal control gene. Ankrd27, Ubc, Tbp, and Actb were consequently used to investigate the effects of internal control genes with differing stabilities on the normalization of target gene expression. Target gene expression levels and changes over time were similar when Ankrd27, Ubc, and Tbp were used as internal controls but different when Actb was used as an internal control. We recommend that Ankrd27, Ubc, and Tbp are used as internal control genes for real-time reverse transcription polymerase chain reaction in spinal cord injury research. This study was approved by the Administration Committee of Experimental Animals, Jiangsu Province, China (approval No. 20180304-008) on March 4, 2018.Wei LiuJie YuYi-Fan WangQian-Qian ShanYa-Xian WangWolters Kluwer Medknow Publicationsarticlegenorm analysis; reference genes; internal control genes; normalization; normfinder analysis; reverse transcription-quantitative polymerase chain reaction; spinal cord injury; stability of gene expressionNeurology. Diseases of the nervous systemRC346-429ENNeural Regeneration Research, Vol 17, Iss 6, Pp 1387-1392 (2022)
institution	DOAJ
collection	DOAJ
language	EN
topic	genorm analysis; reference genes; internal control genes; normalization; normfinder analysis; reverse transcription-quantitative polymerase chain reaction; spinal cord injury; stability of gene expression Neurology. Diseases of the nervous system RC346-429
spellingShingle	genorm analysis; reference genes; internal control genes; normalization; normfinder analysis; reverse transcription-quantitative polymerase chain reaction; spinal cord injury; stability of gene expression Neurology. Diseases of the nervous system RC346-429 Wei Liu Jie Yu Yi-Fan Wang Qian-Qian Shan Ya-Xian Wang Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury
description	There is a lack of systematic research on the expression of internal control genes used for gene expression normalization in real-time reverse transcription polymerase chain reaction in spinal cord injury research. In this study, we used rat models of spinal cord hemisection to analyze the expression stability of 13 commonly applied reference genes: Actb, Ankrd27, CypA, Gapdh, Hprt1, Mrpl10, Pgk1, Rictor, Rn18s, Tbp, Ubc, Ubxn11, and Ywhaz. Our results show that the expression of Ankrd27, Ubc, and Tbp were stable after spinal cord injury, while Actb was the most unstable internal control gene. Ankrd27, Ubc, Tbp, and Actb were consequently used to investigate the effects of internal control genes with differing stabilities on the normalization of target gene expression. Target gene expression levels and changes over time were similar when Ankrd27, Ubc, and Tbp were used as internal controls but different when Actb was used as an internal control. We recommend that Ankrd27, Ubc, and Tbp are used as internal control genes for real-time reverse transcription polymerase chain reaction in spinal cord injury research. This study was approved by the Administration Committee of Experimental Animals, Jiangsu Province, China (approval No. 20180304-008) on March 4, 2018.
format	article
author	Wei Liu Jie Yu Yi-Fan Wang Qian-Qian Shan Ya-Xian Wang
author_facet	Wei Liu Jie Yu Yi-Fan Wang Qian-Qian Shan Ya-Xian Wang
author_sort	Wei Liu
title	Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury
title_short	Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury
title_full	Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury
title_fullStr	Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury
title_full_unstemmed	Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury
title_sort	selection of suitable internal controls for gene expression normalization in rats with spinal cord injury
publisher	Wolters Kluwer Medknow Publications
publishDate	2022
url	https://doaj.org/article/2a51440108954b5f86995299df2515a5
work_keys_str_mv	AT weiliu selectionofsuitableinternalcontrolsforgeneexpressionnormalizationinratswithspinalcordinjury AT jieyu selectionofsuitableinternalcontrolsforgeneexpressionnormalizationinratswithspinalcordinjury AT yifanwang selectionofsuitableinternalcontrolsforgeneexpressionnormalizationinratswithspinalcordinjury AT qianqianshan selectionofsuitableinternalcontrolsforgeneexpressionnormalizationinratswithspinalcordinjury AT yaxianwang selectionofsuitableinternalcontrolsforgeneexpressionnormalizationinratswithspinalcordinjury
_version_	1718420061289447424

Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury

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