Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)

Abstract BMPs are multifunctional growth factors implicated in regulating the ovarian function as key intra-ovarian factors. Biological effects of BMPs are mediated through binding with membrane bound receptors like BMPR-IB and initiating downstream Smad signaling pathway. FecB mutation, regarded as...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sai Kumar, Meeti Punetha, Bosco Jose, Jaya Bharati, Shivani Khanna, Arvind Sonwane, Jonathan A. Green, Kristin Whitworth, Mihir Sarkar
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8d0fd6a6a20d452fa224ad5f322251dc
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:8d0fd6a6a20d452fa224ad5f322251dc
record_format	dspace
spelling	oai:doaj.org-article:8d0fd6a6a20d452fa224ad5f322251dc2021-12-02T12:34:17ZModulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)10.1038/s41598-020-77596-92045-2322https://doaj.org/article/8d0fd6a6a20d452fa224ad5f322251dc2020-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-77596-9https://doaj.org/toc/2045-2322Abstract BMPs are multifunctional growth factors implicated in regulating the ovarian function as key intra-ovarian factors. Biological effects of BMPs are mediated through binding with membrane bound receptors like BMPR-IB and initiating downstream Smad signaling pathway. FecB mutation, regarded as a loss of function mutation in the BMPR-IB gene was identified in certain sheep breeds having high fecundity. Similar type of fecundity genes in goats have not been discovered so far. Hence, the current study was designed to investigate the effects of BMPR-IB gene modulation on granulosa cell function in goats. The BMPR-IB gene was knocked out using CRISPR-Cas technology in granulosa cells and cultured in vitro with BMP-4 stimulation for three different durations In addition, the FecB mutation was introduced in the BMPR-IB gene applying Easi-CRISPR followed by BMP-4/7 stimulation for 72 h. Steroidogenesis and cell viability were studied to explore the granulosa cell function on BMPR-IB gene modulation. BMPRs were found to be expressed stage specifically in granulosa cells of goats. Higher transcriptional abundance of R-Smads, LHR and FSHR indicating sensitisation of Smad signaling and increased gonadotropin sensitivity along with a significant reduction in the cell proliferation and viability was observed in granulosa cells upon BMPR-IB modulation. The inhibitory action of BMP-4/7 on P4 secretion was abolished in both KO and KI cells. Altogether, the study has revealed an altered Smad signaling, steroidogenesis and cell viability upon modulation of BMPR-IB gene in granulosa cells similar to that are documented in sheep breeds carrying the FecB mutation.Sai KumarMeeti PunethaBosco JoseJaya BharatiShivani KhannaArvind SonwaneJonathan A. GreenKristin WhitworthMihir SarkarNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-17 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sai Kumar Meeti Punetha Bosco Jose Jaya Bharati Shivani Khanna Arvind Sonwane Jonathan A. Green Kristin Whitworth Mihir Sarkar Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)
description	Abstract BMPs are multifunctional growth factors implicated in regulating the ovarian function as key intra-ovarian factors. Biological effects of BMPs are mediated through binding with membrane bound receptors like BMPR-IB and initiating downstream Smad signaling pathway. FecB mutation, regarded as a loss of function mutation in the BMPR-IB gene was identified in certain sheep breeds having high fecundity. Similar type of fecundity genes in goats have not been discovered so far. Hence, the current study was designed to investigate the effects of BMPR-IB gene modulation on granulosa cell function in goats. The BMPR-IB gene was knocked out using CRISPR-Cas technology in granulosa cells and cultured in vitro with BMP-4 stimulation for three different durations In addition, the FecB mutation was introduced in the BMPR-IB gene applying Easi-CRISPR followed by BMP-4/7 stimulation for 72 h. Steroidogenesis and cell viability were studied to explore the granulosa cell function on BMPR-IB gene modulation. BMPRs were found to be expressed stage specifically in granulosa cells of goats. Higher transcriptional abundance of R-Smads, LHR and FSHR indicating sensitisation of Smad signaling and increased gonadotropin sensitivity along with a significant reduction in the cell proliferation and viability was observed in granulosa cells upon BMPR-IB modulation. The inhibitory action of BMP-4/7 on P4 secretion was abolished in both KO and KI cells. Altogether, the study has revealed an altered Smad signaling, steroidogenesis and cell viability upon modulation of BMPR-IB gene in granulosa cells similar to that are documented in sheep breeds carrying the FecB mutation.
format	article
author	Sai Kumar Meeti Punetha Bosco Jose Jaya Bharati Shivani Khanna Arvind Sonwane Jonathan A. Green Kristin Whitworth Mihir Sarkar
author_facet	Sai Kumar Meeti Punetha Bosco Jose Jaya Bharati Shivani Khanna Arvind Sonwane Jonathan A. Green Kristin Whitworth Mihir Sarkar
author_sort	Sai Kumar
title	Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)
title_short	Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)
title_full	Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)
title_fullStr	Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)
title_full_unstemmed	Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)
title_sort	modulation of granulosa cell function via crispr-cas fuelled editing of bmpr-ib gene in goats (capra hircus)
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/8d0fd6a6a20d452fa224ad5f322251dc
work_keys_str_mv	AT saikumar modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT meetipunetha modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT boscojose modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT jayabharati modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT shivanikhanna modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT arvindsonwane modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT jonathanagreen modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT kristinwhitworth modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus AT mihirsarkar modulationofgranulosacellfunctionviacrisprcasfuellededitingofbmpribgeneingoatscaprahircus
_version_	1718393870938537984

Modulation of granulosa cell function via CRISPR-Cas fuelled editing of BMPR-IB gene in goats (Capra hircus)

Ejemplares similares