CRISPR/Cas9 mediated approach to generate YAP-depleted human embryonic stem cell line (MUSIe002-A-1)

Khampang, Sujittra; Jiamvoraphong, Nittaya; Laowtammathron, Chuti; Lorthongpanich, Chanchao; Klaihmon, Phatchanat; Srisook, Pimonwan; Kang, Xin; Luanpitpong, Sudjit; U-Pratya, Yaowalak; Issaragrisil, Surapol

doi:10.1016/j.scr.2022.102990

Cited by 1 publication

(2 citation statements)

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“…To investigate the role of YAP during human spermatogenesis, the isogenic YAP-depleted hESCs (YAP-KD cells) were established by transducing hESCs (MUSIe002-A cells) with the YAP -targeted CRISPR/Cas9 plasmid 33 . Western blot analysis showed a 50% reduction in the YAP level in YAP-KD cells compared to their wild-type control (WT cells) (Fig.…”

Section: Resultsmentioning

confidence: 99%

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The dynamic expression of YAP is essential for the development of male germ cells derived from human embryonic stem cells

Khampang,

Lorthongpanich,

Laowtammathron

et al. 2024

Sci Rep

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YAP plays a vital role in controlling growth and differentiation in various cell lineages. Although the expression of YAP in mice testicular and spermatogenic cells suggests its role in mammalian spermatogenesis, the role of YAP in the development of human male germ cells has not yet been determined. Using an in vitro model and a gene editing approach, we generated human spermatogonia stem cell-like cells (hSSLCs) from human embryonic stem cells (hESCs) and investigated the role of YAP in human spermatogenesis. The results showed that reducing YAP expression during the early stage of spermatogenic differentiation increased the number of PLZF+ hSSLCs and haploid spermatid-like cells. We also demonstrated that the up-regulation of YAP is essential for maintaining spermatogenic cell survival during the later stages of spermatogenic differentiation. The expression of YAP that deviates from this pattern results in a lower number of hSSLCs and an increased level of spermatogenic cell death. Taken together, our result demonstrates that the dynamic expression pattern of YAP is essential for human spermatogenesis. Modulating the level of YAP during human spermatogenesis could improve the production yield of male germ cells derived from hESCs, which could provide the optimization method for in vitro gametogenesis and gain insight into the application in the treatment of male infertility.

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Section: Resultsmentioning

confidence: 99%

“…After 5–7 days of culture, the emerging colonies were picked up, expanded, and harvested to determine their YAP expression by Western Blot. The YAP knockdown hESCs, MUSIe002-A-1, were successfully established 33 and later referred to as YAP-KD cells in this manuscript.…”

Section: Methodsmentioning

confidence: 99%