Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY Gonad

Delivering siRNA or shRNA into the developing embryos is still a main challenge to use of RNAi in mammalian systems. Here we analyze several factors influencing RNAi-mediated silencing of Sry gene, which is a tightly controlled spatiotemporal expressed gene and only shortly expressed in developing m...

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Main Authors: Ning Wu, Ai-Bing Yu, Hua-Bin Zhu, Xiu-Kun Lin
Format: Article
Language:English
Published: Hindawi Limited 2012-01-01
Series:Journal of Biomedicine and Biotechnology
Online Access:http://dx.doi.org/10.1155/2012/343891
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spelling doaj-a0d7b53c98dd4050a2c5557181ab9a682020-11-25T01:57:47ZengHindawi LimitedJournal of Biomedicine and Biotechnology1110-72431110-72512012-01-01201210.1155/2012/343891343891Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY GonadNing Wu0Ai-Bing Yu1Hua-Bin Zhu2Xiu-Kun Lin3Institute of Oceanology, Chinese Academy of Sciences, No. 7 of Nanhai Road, Qingdao 266071, ChinaLaboratory of Animal Nutrition, Graduate School of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaInstitute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, ChinaInstitute of Oceanology, Chinese Academy of Sciences, No. 7 of Nanhai Road, Qingdao 266071, ChinaDelivering siRNA or shRNA into the developing embryos is still a main challenge to use of RNAi in mammalian systems. Here we analyze several factors influencing RNAi-mediated silencing of Sry gene, which is a tightly controlled spatiotemporal expressed gene and only shortly expressed in developing mouse embryo gonad. A Sry gene-specific shRNAs expression vector (pSilencer4.1/Sry565) was constructed. The shRNA constructs were mixed with polyethylenimines (PEIs) to form a complex and then injected into pregnant mice though tail vein. Our results showed that Sry gene was downregulated significantly in developing embryos. Further study revealed that knocking-down of Sry expression resulted in feminization of gonad development in mouse embryos and the expression level of Sox9 and Wt1 gene was also significantly changed by downregulation of Sry. The transfection efficiency is associated with the amount of plasmid DNA injection, injection time, injection speed, and volume. Our studies suggest that transplacental RNAi could be implemented by tail vein injection of plasmid vector into pregnant mice.http://dx.doi.org/10.1155/2012/343891
collection DOAJ
language English
format Article
sources DOAJ
author Ning Wu
Ai-Bing Yu
Hua-Bin Zhu
Xiu-Kun Lin
spellingShingle Ning Wu
Ai-Bing Yu
Hua-Bin Zhu
Xiu-Kun Lin
Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY Gonad
Journal of Biomedicine and Biotechnology
author_facet Ning Wu
Ai-Bing Yu
Hua-Bin Zhu
Xiu-Kun Lin
author_sort Ning Wu
title Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY Gonad
title_short Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY Gonad
title_full Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY Gonad
title_fullStr Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY Gonad
title_full_unstemmed Effective Silencing of Sry Gene with RNA Interference in Developing Mouse Embryos Resulted in Feminization of XY Gonad
title_sort effective silencing of sry gene with rna interference in developing mouse embryos resulted in feminization of xy gonad
publisher Hindawi Limited
series Journal of Biomedicine and Biotechnology
issn 1110-7243
1110-7251
publishDate 2012-01-01
description Delivering siRNA or shRNA into the developing embryos is still a main challenge to use of RNAi in mammalian systems. Here we analyze several factors influencing RNAi-mediated silencing of Sry gene, which is a tightly controlled spatiotemporal expressed gene and only shortly expressed in developing mouse embryo gonad. A Sry gene-specific shRNAs expression vector (pSilencer4.1/Sry565) was constructed. The shRNA constructs were mixed with polyethylenimines (PEIs) to form a complex and then injected into pregnant mice though tail vein. Our results showed that Sry gene was downregulated significantly in developing embryos. Further study revealed that knocking-down of Sry expression resulted in feminization of gonad development in mouse embryos and the expression level of Sox9 and Wt1 gene was also significantly changed by downregulation of Sry. The transfection efficiency is associated with the amount of plasmid DNA injection, injection time, injection speed, and volume. Our studies suggest that transplacental RNAi could be implemented by tail vein injection of plasmid vector into pregnant mice.
url http://dx.doi.org/10.1155/2012/343891
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