A high‐throughput screen in mESCs to identify the cross‐talk between signaling, endocytosis, and pluripotency.
| Title: | A high‐throughput screen in mESCs to identify the cross‐talk between signaling, endocytosis, and pluripotency. |
|---|---|
| Authors: | Mote, Ridim D.1 (AUTHOR), Tiwari, Mahak1,2 (AUTHOR), Yadavalli, Narayana1 (AUTHOR), Rajan, Raghav3 (AUTHOR), Subramanyam, Deepa1 (AUTHOR) deepa@nccs.res.in |
| Source: | Cell Biology International. Jul2024, Vol. 48 Issue 7, p1035-1046. 12p. |
| Subject Terms: | *EMBRYONIC stem cells, *HIGH throughput screening (Drug development), *ENDOCYTOSIS, *SOX2 protein, *GREEN fluorescent protein, *COATED vesicles, *STEM cells |
| Reviews & Products: | MATLAB (Computer software) |
| Abstract: | Embryonic stem cell fate is regulated by various cellular processes. Recently, the process of endocytosis has been implicated in playing a role in the maintenance of self‐renewal and pluripotency of mouse embryonic stem cells. A previous siRNA‐based screen interrogated the function of core components of the endocytic machinery in maintaining the pluripotency of embryonic stem cells, revealing a crucial role for clathrin mediated endocytosis. A number of other proteins involved in key signaling pathways have also been shown to both regulate and be regulated by endocytosis. We collated a list of 1141 genes in connection to the term "endocytosis" from Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO), excluding those previously interrogated, and examined the effect of their knockdown on the pluripotency of mouse embryonic stem cells (mESCs) using levels of green fluorescent protein driven by the Oct4 promoter. We used high‐throughput screening followed by an automated MATrix LABoratory (MATLAB)‐based analysis pipeline and assessed changes in GFP fluorescence as a readout for ESC pluripotency. Through this screen we identified a number of genes, many hitherto not associated with stem cell pluripotency, which upon knockdown either resulted in a significant increase or decrease of GFP fluorescence. We further present validation for some of these hits. We present a workflow aimed to identify genes involved in signaling pathways which can be regulated by endocytosis, and that affect the pluripotency of ESCs. [ABSTRACT FROM AUTHOR] |
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| Database: | Academic Search Complete |
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| Items | – Name: Title Label: Title Group: Ti Data: A high‐throughput screen in mESCs to identify the cross‐talk between signaling, endocytosis, and pluripotency. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Mote%2C+Ridim+D%2E%22">Mote, Ridim D.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Tiwari%2C+Mahak%22">Tiwari, Mahak</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Yadavalli%2C+Narayana%22">Yadavalli, Narayana</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Rajan%2C+Raghav%22">Rajan, Raghav</searchLink><relatesTo>3</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Subramanyam%2C+Deepa%22">Subramanyam, Deepa</searchLink><relatesTo>1</relatesTo> (AUTHOR)<i> deepa@nccs.res.in</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Cell+Biology+International%22">Cell Biology International</searchLink>. Jul2024, Vol. 48 Issue 7, p1035-1046. 12p. – Name: Subject Label: Subject Terms Group: Su Data: *<searchLink fieldCode="DE" term="%22EMBRYONIC+stem+cells%22">EMBRYONIC stem cells</searchLink><br />*<searchLink fieldCode="DE" term="%22HIGH+throughput+screening+%28Drug+development%29%22">HIGH throughput screening (Drug development)</searchLink><br />*<searchLink fieldCode="DE" term="%22ENDOCYTOSIS%22">ENDOCYTOSIS</searchLink><br />*<searchLink fieldCode="DE" term="%22SOX2+protein%22">SOX2 protein</searchLink><br />*<searchLink fieldCode="DE" term="%22GREEN+fluorescent+protein%22">GREEN fluorescent protein</searchLink><br />*<searchLink fieldCode="DE" term="%22COATED+vesicles%22">COATED vesicles</searchLink><br />*<searchLink fieldCode="DE" term="%22STEM+cells%22">STEM cells</searchLink> – Name: SubjectProduct Label: Reviews & Products Group: Su Data: <searchLink fieldCode="PS" term="%22MATLAB+%28Computer+software%29%22">MATLAB (Computer software)</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Embryonic stem cell fate is regulated by various cellular processes. Recently, the process of endocytosis has been implicated in playing a role in the maintenance of self‐renewal and pluripotency of mouse embryonic stem cells. A previous siRNA‐based screen interrogated the function of core components of the endocytic machinery in maintaining the pluripotency of embryonic stem cells, revealing a crucial role for clathrin mediated endocytosis. A number of other proteins involved in key signaling pathways have also been shown to both regulate and be regulated by endocytosis. We collated a list of 1141 genes in connection to the term "endocytosis" from Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO), excluding those previously interrogated, and examined the effect of their knockdown on the pluripotency of mouse embryonic stem cells (mESCs) using levels of green fluorescent protein driven by the Oct4 promoter. We used high‐throughput screening followed by an automated MATrix LABoratory (MATLAB)‐based analysis pipeline and assessed changes in GFP fluorescence as a readout for ESC pluripotency. Through this screen we identified a number of genes, many hitherto not associated with stem cell pluripotency, which upon knockdown either resulted in a significant increase or decrease of GFP fluorescence. We further present validation for some of these hits. We present a workflow aimed to identify genes involved in signaling pathways which can be regulated by endocytosis, and that affect the pluripotency of ESCs. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Cell Biology International is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1002/cbin.12168 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 12 StartPage: 1035 Subjects: – SubjectFull: MATLAB (Computer software) Type: general – SubjectFull: EMBRYONIC stem cells Type: general – SubjectFull: HIGH throughput screening (Drug development) Type: general – SubjectFull: ENDOCYTOSIS Type: general – SubjectFull: SOX2 protein Type: general – SubjectFull: GREEN fluorescent protein Type: general – SubjectFull: COATED vesicles Type: general – SubjectFull: STEM cells Type: general Titles: – TitleFull: A high‐throughput screen in mESCs to identify the cross‐talk between signaling, endocytosis, and pluripotency. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Mote, Ridim D. – PersonEntity: Name: NameFull: Tiwari, Mahak – PersonEntity: Name: NameFull: Yadavalli, Narayana – PersonEntity: Name: NameFull: Rajan, Raghav – PersonEntity: Name: NameFull: Subramanyam, Deepa IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 07 Text: Jul2024 Type: published Y: 2024 Identifiers: – Type: issn-print Value: 10656995 Numbering: – Type: volume Value: 48 – Type: issue Value: 7 Titles: – TitleFull: Cell Biology International Type: main |
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