Application of a novel cell-permeable peptide-driven protein delivery in mouse blastocysts

Kwon, Sunghoon; Kwak, Areum; Shin, Hyejin; Choi, Sie−Young; Kim, Soohyun; Lim, Hyunjung Jade

doi:10.1530/rep-13-0203

Cited by 7 publications

(4 citation statements)

References 31 publications

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“…Previous studies have demonstrated that hydrophilic arginine-rich cell-penetrating peptides have optimal internalization capability for small molecules in mammalian cells (Mitchell et al, 2000;Mueller et al, 2008;Tünnemann et al, 2008). Presently, there is a limited amount of literature available on the utilization of CPPs in oocytes (Kwon et al, 2009;Kwon et al, 2013;Yang et al, 2014;Jeon et al, 2019;Zhu et al, 2021), and none of these studies extensively examine the internalization of CPPs into mouse oocytes. First, we evaluated the ability of CPPs to cross both the zona pellucida and plasma membrane of immature (GV) and mature (MII) mouse oocytes within a 1 h incubation period.…”

Section: Discussionmentioning

confidence: 99%

In Celebration of Women in Molecular and Cellular Reproduction

2024

Frontiers Research Topics

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Frontiers is more than just an open access publisher of scholarly articles: it is a pioneering approach to the world of academia, radically improving the way scholarly research is managed. The grand vision of Frontiers is a world where all people have an equal opportunity to seek, share and generate knowledge. Frontiers provides immediate and permanent online open access to all its publications, but this alone is not enough to realize our grand goals. Frontiers journal seriesThe Frontiers journal series is a multi-tier and interdisciplinary set of openaccess, online journals, promising a paradigm shift from the current review, selection and dissemination processes in academic publishing. All Frontiers journals are driven by researchers for researchers; therefore, they constitute a service to the scholarly community. At the same time, the Frontiers journal series operates on a revolutionary invention, the tiered publishing system, initially addressing specific communities of scholars, and gradually climbing up to broader public understanding, thus serving the interests of the lay society, too. Dedication to qualityEach Frontiers article is a landmark of the highest quality, thanks to genuinely collaborative interactions between authors and review editors, who include some of the world's best academicians. Research must be certified by peers before entering a stream of knowledge that may eventually reach the public -and shape society; therefore, Frontiers only applies the most rigorous and unbiased reviews. Frontiers revolutionizes research publishing by freely delivering the most outstanding research, evaluated with no bias from both the academic and social point of view. By applying the most advanced information technologies, Frontiers is catapulting scholarly publishing into a new generation. What are Frontiers Research Topics?Frontiers Research Topics are very popular trademarks of the Frontiers journals series: they are collections of at least ten articles, all centered on a particular subject. With their unique mix of varied contributions from Original Research to Review Articles, Frontiers Research Topics unify the most influential researchers, the latest key findings and historical advances in a hot research area.

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Section: Discussionmentioning

confidence: 99%

In Celebration of Women in Molecular and Cellular Reproduction

2024

Frontiers Research Topics

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“…Previous studies have demonstrated that hydrophilic arginine-rich cell-penetrating peptides have optimal internalization capability for small molecules in mammalian cells ( Mitchell et al, 2000 ; Mueller et al, 2008 ; Tünnemann et al, 2008 ). Presently, there is a limited amount of literature available on the utilization of CPPs in oocytes ( Kwon et al, 2009 ; Kwon et al, 2013 ; Yang et al, 2014 ; Jeon et al, 2019 ; Zhu et al, 2021 ), and none of these studies extensively examine the internalization of CPPs into mouse oocytes.…”

Section: Discussionmentioning

confidence: 99%

“…The same group also tested a 12-mer CPP from the conserved region of the human papillomavirus L1 capsid protein, LDP12. Nevertheless, CPP was not internalized in mature oocytes (with or without ZP) and was not assayed in immature oocytes (S. Kwon et al, 2013 ). In this work, we have demonstrated that CPP internalization observation depends on the mounting method.…”

Section: Discussionmentioning

confidence: 99%

The light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibits cortical reaction in mouse oocytes

Klinsky,

Wetten,

Zanni-Ruiz

et al. 2023

Front. Cell Dev. Biol.

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Introduction: Cortical reaction is a secretory process that occurs after a spermatozoon fuses with the oocyte, avoiding the fusion of additional sperm. During this exocytic event, the cortical granule membrane fuses with the oocyte plasma membrane. We have identified several molecular components involved in this process and confirmed that SNARE proteins regulate membrane fusion during cortical reaction in mouse oocytes. In those studies, we microinjected different nonpermeable reagents to demonstrate the participation of a specific protein in the cortical reaction. However, the microinjection technique has several limitations. In this work, we aimed to assess the potential of cell-penetrating peptides (CPP) as biotechnological tools for delivering molecules into oocytes, and to evaluate the functionality of the permeable tetanus toxin (bound to CPP sequence) during cortical reaction.Methods: Arginine-rich cell-penetrating peptides have demonstrated the optimal internalization of small molecules in mammalian cells. Two arginine-rich CPP were used in the present study. One, labeled with 5-carboxyfluorescein, to characterize the factors that can modulate its internalization, and the other, the permeable light chain of tetanus toxin, that cleaves the SNAREs VAMP1 and VAMP3 expressed in mouse oocytes.Results: Results showed that fluorescent CPP was internalized into the oocyte cytoplasm and that internalization was dependent on the concentration, time, temperature, and maturation stage of the oocyte. Using our functional assay to study cortical reaction, the light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibited cortical granules exocytosis.Discussion: Results obtained from the use of permeable peptides demonstrate that this CPP is a promising biotechnological tool to study functional macromolecules in mouse oocytes.

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“…It was recently reported that induced pluripotent cells (iPSCs) could be generated by introducing transcription factors fused with a cell-penetrating peptide (CPP) 21 , 22 . Lim and colleagues also identified a new form of CPP, obtained from human papillomavirus L1 capsid protein (LDP12), and confirmed its ability to deliver a fusion protein of enhanced green fluorescence protein and MAP1LC3 (EGFP-LC3) into mouse blastocysts 23 . In our previously reports, the cell number of inner cell mass (ICM) and expression of the Oct4 gene were positively regulated in fresh and cryopreserved embryos after treatment with CPP-conjugated estrogen-related receptor β (CPP-ESRRB) during in vitro cultivation 24 , 25 .…”

Section: Introductionmentioning

confidence: 92%

The effect of cell penetrating peptide-conjugated coactivator-associated arginine methyltransferase 1 (CPP-CARM1) on the cloned mouse embryonic development

Bang

Lee

et al. 2018

Sci Rep

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Abnormalities in gene expression that negatively affect embryonic development are frequently observed in cloned embryos generated by somatic cell nuclear transfer (SCNT). In the present study, we successfully produced a cell-penetrating peptide (CPP)-conjugated with coactivator-associated arginine methyltransferase 1 (CARM1) protein from mammalian cells and confirmed introduction into donor somatic cells and cloned 8-cell embryos within 3 hours after addition to culture medium. In addition, H3R17 dimethylation and embryonic development up to the blastocyst stage were increased in the group treated with exogenous CPP-CARM1 protein compared with the untreated group (control). Interestingly, the number of total cells and trophectoderm in blastocysts as well as implantation rate were significantly increased in the CPP-CARM1 protein-treated group. However, the cell number of inner cell mass (ICM) was not changed compared with the control group; similarly, expression of pluripotency-related genes Oct4 and Nanog (ICM markers) was not significantly different between groups. On the other hand, expression of the implantation-related gene Cdx2 (trophectoderm marker) was transiently increased after treatment with CPP-CARM1 protein. On the basis of these results, we conclude that supplementation with exogenous CPP-CARM1 protein improves embryonic development of cloned embryos through regulation of histone methylation and gene expression. In addition, our results suggest that CPP-CARM1 protein may be a useful tool for strengthening implantation of mammalian embryos.

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Application of a novel cell-permeable peptide-driven protein delivery in mouse blastocysts

Cited by 7 publications

References 31 publications

In Celebration of Women in Molecular and Cellular Reproduction

In Celebration of Women in Molecular and Cellular Reproduction

The light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibits cortical reaction in mouse oocytes

The effect of cell penetrating peptide-conjugated coactivator-associated arginine methyltransferase 1 (CPP-CARM1) on the cloned mouse embryonic development

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