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Objectives <p>Gene therapy has emerged as an encouraging therapeutic approach for several diseases. The use of plasmid DNA (pDNA)&#xa0;or viral vectors encoding proteins have yielded low efficacy in recent trials. Synthetic&#xa0;messenger RNA&#xa0;(mRNA), also known as <i>in vitro</i> transcribed mRNA (IVT-mRNA), is a highly potent alternative. However, the use of IVT-mRNA in reproductive conditions remains scarce. Our previous work has demonstrated the involvement of endometrial serum- and glucocorticoid kinase 1 (SGK1), where it influences pregnancy outcomes in murine models. On this basis, we investigated the expression and function of human SGK1 encoded by IVT-mRNA <i>in vitro</i>.</p> Results <p>We show using flow cytometry, quantitative RT-PCR and immunoblotting, that IVT-mRNA has a superior transfection efficiency compared with pDNA. At protein level, we show that the constitutively active SGK1 variant (<i>SGK1</i><sub>S422D</sub>) could increase protein levels and&#xa0;that the inactive SGK1 variant (<i>SGK1</i><sub>K127A</sub>) could reduce SGK1 levels. This effect of SGK1 activity modulation could contribute to reprogramming endometrial cells and support a conducive environment for pregnancy.</p>

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Optimizing the use of in vitro transcribed SGK1-mRNA as a therapeutic tool to treat female infertility

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年7月23日

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  • Itishri Sahu,
  • Fabienne Engelmann,
  • Brian Weidensee,
  • Harivignesh Ganesan,
  • Sara Y. Brucker,
  • Yogesh Singh,
  • Madhuri S. Salker

摘要

Objectives

Gene therapy has emerged as an encouraging therapeutic approach for several diseases. The use of plasmid DNA (pDNA) or viral vectors encoding proteins have yielded low efficacy in recent trials. Synthetic messenger RNA (mRNA), also known as in vitro transcribed mRNA (IVT-mRNA), is a highly potent alternative. However, the use of IVT-mRNA in reproductive conditions remains scarce. Our previous work has demonstrated the involvement of endometrial serum- and glucocorticoid kinase 1 (SGK1), where it influences pregnancy outcomes in murine models. On this basis, we investigated the expression and function of human SGK1 encoded by IVT-mRNA in vitro.

Results

We show using flow cytometry, quantitative RT-PCR and immunoblotting, that IVT-mRNA has a superior transfection efficiency compared with pDNA. At protein level, we show that the constitutively active SGK1 variant (SGK1S422D) could increase protein levels and that the inactive SGK1 variant (SGK1K127A) could reduce SGK1 levels. This effect of SGK1 activity modulation could contribute to reprogramming endometrial cells and support a conducive environment for pregnancy.