Comparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in mice

The Gt(ROSA)26Sor (ROSA26) and H11 loci are commonly used as safe harbors for the construction of targeted transgenic mice. However, it remains unclear whether these two loci have distinct effects on transgene expression. In this study, we insert three differently colored fluorescent protein express...

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Main Authors: Yu Zhilan, Shi Jiahao, Zhang Jingyu, Wu Youbing, Shen Ruling, Fei Jian
Format: Article
Language:English
Published: China Science Publishing & Media Ltd. 2024-05-01
Series:Acta Biochimica et Biophysica Sinica
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Online Access:https://www.sciengine.com/doi/10.3724/abbs.2024081
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author Yu Zhilan
Shi Jiahao
Zhang Jingyu
Wu Youbing
Shen Ruling
Fei Jian
author_facet Yu Zhilan
Shi Jiahao
Zhang Jingyu
Wu Youbing
Shen Ruling
Fei Jian
author_sort Yu Zhilan
collection DOAJ
description The Gt(ROSA)26Sor (ROSA26) and H11 loci are commonly used as safe harbors for the construction of targeted transgenic mice. However, it remains unclear whether these two loci have distinct effects on transgene expression. In this study, we insert three differently colored fluorescent protein expression cassettes (EGFP, tdTomato and mTagBFP2) driven by the CAG promoter into the ROSA26 and H11 loci. We generate five single-transgenic mouse models and a triple-transgenic mouse model expressing three distinct fluorescent proteins simultaneously. Our results reveal that the efficiency of transgene expression is greater at the ROSA26 locus with a reverse orientation relative to the transcription of the ROSA26 gene. In most tissues examined, the efficiency of transgene expression at the ROSA26 locus exceeds that at the H11 locus. Moreover, the expression profiles of identical transgenes display discrepancies across various tissues, and notably, substantial heterogeneity in transgene expression is discernible within cells of the same tissue. Our findings offer a valuable reference for the selection of safe harbors and strategies for the construction of transgenic mouse models.
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spelling doaj-art-1701b06e25594f5d9393c647733bc2f12025-08-20T02:32:38ZengChina Science Publishing & Media Ltd.Acta Biochimica et Biophysica Sinica1672-91452024-05-01561687169810.3724/abbs.202408120d259ccComparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in miceYu Zhilan0Shi Jiahao1Zhang Jingyu2Wu Youbing3Shen Ruling4Fei Jian5["School of Life Science and Technology, Tongji University, Shanghai 200092, China"]["School of Life Science and Technology, Tongji University, Shanghai 200092, China"]["School of Life Science and Technology, Tongji University, Shanghai 200092, China"]["Shanghai Engineering Research Center for Model Organisms, SMOC, Shanghai 201203, China"]["Shanghai Laboratory Animal Research Center, Shanghai 201203, China"]["School of Life Science and Technology, Tongji University, Shanghai 200092, China","Shanghai Engineering Research Center for Model Organisms, SMOC, Shanghai 201203, China"]The Gt(ROSA)26Sor (ROSA26) and H11 loci are commonly used as safe harbors for the construction of targeted transgenic mice. However, it remains unclear whether these two loci have distinct effects on transgene expression. In this study, we insert three differently colored fluorescent protein expression cassettes (EGFP, tdTomato and mTagBFP2) driven by the CAG promoter into the ROSA26 and H11 loci. We generate five single-transgenic mouse models and a triple-transgenic mouse model expressing three distinct fluorescent proteins simultaneously. Our results reveal that the efficiency of transgene expression is greater at the ROSA26 locus with a reverse orientation relative to the transcription of the ROSA26 gene. In most tissues examined, the efficiency of transgene expression at the ROSA26 locus exceeds that at the H11 locus. Moreover, the expression profiles of identical transgenes display discrepancies across various tissues, and notably, substantial heterogeneity in transgene expression is discernible within cells of the same tissue. Our findings offer a valuable reference for the selection of safe harbors and strategies for the construction of transgenic mouse models.https://www.sciengine.com/doi/10.3724/abbs.2024081transgenic mouseknockin<italic>Gt(ROSA)26Sor</italic><italic>H11</italic>gene expression
spellingShingle Yu Zhilan
Shi Jiahao
Zhang Jingyu
Wu Youbing
Shen Ruling
Fei Jian
Comparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in mice
Acta Biochimica et Biophysica Sinica
transgenic mouse
knockin
<italic>Gt(ROSA)26Sor</italic>
<italic>H11</italic>
gene expression
title Comparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in mice
title_full Comparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in mice
title_fullStr Comparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in mice
title_full_unstemmed Comparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in mice
title_short Comparative study on the expression characteristics of transgenes inserted into the Gt(ROSA)26Sor and H11 loci in mice
title_sort comparative study on the expression characteristics of transgenes inserted into the gt rosa 26sor and h11 loci in mice
topic transgenic mouse
knockin
<italic>Gt(ROSA)26Sor</italic>
<italic>H11</italic>
gene expression
url https://www.sciengine.com/doi/10.3724/abbs.2024081
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AT shijiahao comparativestudyontheexpressioncharacteristicsoftransgenesinsertedintothegtrosa26sorandh11lociinmice
AT zhangjingyu comparativestudyontheexpressioncharacteristicsoftransgenesinsertedintothegtrosa26sorandh11lociinmice
AT wuyoubing comparativestudyontheexpressioncharacteristicsoftransgenesinsertedintothegtrosa26sorandh11lociinmice
AT shenruling comparativestudyontheexpressioncharacteristicsoftransgenesinsertedintothegtrosa26sorandh11lociinmice
AT feijian comparativestudyontheexpressioncharacteristicsoftransgenesinsertedintothegtrosa26sorandh11lociinmice