LIVIVO - Search results -

Search results

Result 1 - 10 of total 17

Search options

Article ; Online: CRISPR-Cas9 screen in human embryonic stem cells to identify genes required for neural differentiation.

Qi, Shutao / Sivakumar, Sushama / Yu, Hongtao

STAR protocols

2022 Volume 3, Issue 4, Page(s) 101682

Abstract: Human embryonic stem cells (hESCs) continuously self-renew in culture and can be induced to differentiate into multiple cell types, including neural progenitor cells (NPCs). Here, we present a protocol to perform a CRISPR-Cas9 screen in hESCs to identify ...

Abstract	Human embryonic stem cells (hESCs) continuously self-renew in culture and can be induced to differentiate into multiple cell types, including neural progenitor cells (NPCs). Here, we present a protocol to perform a CRISPR-Cas9 screen in hESCs to identify regulators that promote SOX1 expression during NPC formation. This screening protocol can be adapted with other endpoint reporters for the identification of genes involved in the commitment of hESCs to other cell lineages. For complete details on the use and execution of this protocol, please refer to Sivakumar et al. (2022).
MeSH term(s)	Humans ; Human Embryonic Stem Cells ; CRISPR-Cas Systems ; Neural Stem Cells ; Cell Differentiation
Language	English
Publishing date	2022-09-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2022.101682
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

Inter-library loan at ZB MED

Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

Article: Phosphatase-regulated recruitment of the spindle- and kinetochore-associated (Ska) complex to kinetochores.

Sivakumar, Sushama / Gorbsky, Gary J

Biology open

2017 Volume 6, Issue 11, Page(s) 1672–1679

Abstract: Kinetochores move chromosomes on dynamic spindle microtubules and regulate signaling of the spindle checkpoint. The spindle- and kinetochore-associated (Ska) complex, a hexamer composed of two copies of Ska1, Ska2 and Ska3, has been implicated in both ... ...

Abstract	Kinetochores move chromosomes on dynamic spindle microtubules and regulate signaling of the spindle checkpoint. The spindle- and kinetochore-associated (Ska) complex, a hexamer composed of two copies of Ska1, Ska2 and Ska3, has been implicated in both roles. Phosphorylation of kinetochore components by the well-studied mitotic kinases Cdk1, Aurora B, Plk1, Mps1, and Bub1 regulate chromosome movement and checkpoint signaling. Roles for the opposing phosphatases are more poorly defined. Recently, we showed that the C terminus of Ska1 recruits protein phosphatase 1 (PP1) to kinetochores. Here we show that PP1 and protein phosphatase 2A (PP2A) both promote accumulation of Ska at kinetochores. Depletion of PP1 or PP2A by siRNA reduces Ska binding at kinetochores, impairs alignment of chromosomes to the spindle midplane, and causes metaphase delay or arrest, phenotypes that are also seen after depletion of Ska. Artificial tethering of PP1 to the outer kinetochore protein Nuf2 promotes Ska recruitment to kinetochores, and it reduces but does not fully rescue chromosome alignment and metaphase arrest defects seen after Ska depletion. We propose that Ska has multiple functions in promoting mitotic progression and that kinetochore-associated phosphatases function in a positive feedback cycle to reinforce Ska complex accumulation at kinetochores.
Language	English
Publishing date	2017-11-15
Publishing country	England
Document type	Journal Article
ZDB-ID	2632264-X
ISSN	2046-6390
ISSN	2046-6390
DOI	10.1242/bio.026930
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article ; Online: CHD-associated enhancers shape human cardiomyocyte lineage commitment.

Armendariz, Daniel A / Goetsch, Sean C / Sundarrajan, Anjana / Sivakumar, Sushama / Wang, Yihan / Xie, Shiqi / Munshi, Nikhil V / Hon, Gary C

eLife

2023 Volume 12

Abstract: Enhancers orchestrate gene expression programs that drive multicellular development and lineage commitment. Thus, genetic variants at enhancers are thought to contribute to developmental diseases by altering cell fate commitment. However, while many ... ...

Abstract	Enhancers orchestrate gene expression programs that drive multicellular development and lineage commitment. Thus, genetic variants at enhancers are thought to contribute to developmental diseases by altering cell fate commitment. However, while many variant-containing enhancers have been identified, studies to endogenously test the impact of these enhancers on lineage commitment have been lacking. We perform a single-cell CRISPRi screen to assess the endogenous roles of 25 enhancers and putative cardiac target genes implicated in genetic studies of congenital heart defects (CHDs). We identify 16 enhancers whose repression leads to deficient differentiation of human cardiomyocytes (CMs). A focused CRISPRi validation screen shows that repression of TBX5 enhancers delays the transcriptional switch from mid- to late-stage CM states. Endogenous genetic deletions of two TBX5 enhancers phenocopy epigenetic perturbations. Together, these results identify critical enhancers of cardiac development and suggest that misregulation of these enhancers could contribute to cardiac defects in human patients.
MeSH term(s)	Humans ; Myocytes, Cardiac/metabolism ; Regulatory Sequences, Nucleic Acid ; Cell Differentiation/genetics ; Heart Defects, Congenital/genetics
Language	English
Publishing date	2023-04-25
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.86206
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article ; Online: Live Fluorescence Imaging of Chromosome Segregation in Cultured Cells.

Daum, John R / DuBose, Casey O / Sivakumar, Sushama / Gorbsky, Gary J

Methods in molecular biology (Clifton, N.J.)

2021 Volume 2415, Page(s) 61–86

Abstract: Live-cell fluorescence microscopy is an effective tool for characterizing aberrant mitotic phenotypes resulting from exposure to chemical inhibitors and after RNA interference-mediated or CRISPR knockout-mediated depletion of protein targets. Live ... ...

Abstract	Live-cell fluorescence microscopy is an effective tool for characterizing aberrant mitotic phenotypes resulting from exposure to chemical inhibitors and after RNA interference-mediated or CRISPR knockout-mediated depletion of protein targets. Live imaging of cultured cells during mitotic progression presents challenges in maintaining optimal health of cells while achieving the temporal and spatial resolution to accomplish the goals of the study. Herein are strategies to monitor and analyze mammalian cell mitosis utilizing either a wide field or a light sheet, inverted, fluorescence microscope.
MeSH term(s)	Cells, Cultured ; Chromosome Segregation ; Microscopy, Fluorescence/methods ; Mitosis ; Optical Imaging
Language	English
Publishing date	2021-12-21
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-1904-9_5
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Inter-library loan at ZB MED

Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

Article ; Online: Spatiotemporal regulation of the anaphase-promoting complex in mitosis.

Sivakumar, Sushama / Gorbsky, Gary J

Nature reviews. Molecular cell biology

2015 Volume 16, Issue 2, Page(s) 82–94

Abstract: The appropriate timing of events that lead to chromosome segregation during mitosis and cytokinesis is essential to prevent aneuploidy, and defects in these processes can contribute to tumorigenesis. Key mitotic regulators are controlled through ... ...

Abstract	The appropriate timing of events that lead to chromosome segregation during mitosis and cytokinesis is essential to prevent aneuploidy, and defects in these processes can contribute to tumorigenesis. Key mitotic regulators are controlled through ubiquitylation and proteasome-mediated degradation. The APC/C (anaphase-promoting complex; also known as the cyclosome) is an E3 ubiquitin ligase that has a crucial function in the regulation of the mitotic cell cycle, particularly at the onset of anaphase and during mitotic exit. Co-activator proteins, inhibitor proteins, protein kinases and phosphatases interact with the APC/C to temporally and spatially control its activity and thus ensure accurate timing of mitotic events.
MeSH term(s)	Anaphase-Promoting Complex-Cyclosome/metabolism ; Cell Cycle Proteins/metabolism ; Chromosome Segregation/physiology ; Cytokinesis/physiology ; Humans ; Mitosis/physiology
Chemical Substances	Cell Cycle Proteins ; Anaphase-Promoting Complex-Cyclosome (EC 2.3.2.27)
Language	English
Publishing date	2015-02
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2031313-5
ISSN	1471-0080 ; 1471-0072
ISSN (online)	1471-0080
ISSN	1471-0072
DOI	10.1038/nrm3934
Database	MEDical Literature Analysis and Retrieval System OnLINE

Full text online

Full text

In stock of ZB MED Cologne/Königswinter

Zs.A 5446: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 26: Show issues

Order via subito

Details ▾

Article ; Online: TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling.

Sivakumar, Sushama / Qi, Shutao / Cheng, Ningyan / Sathe, Adwait A / Kanchwala, Mohammed / Kumar, Ashwani / Evers, Bret M / Xing, Chao / Yu, Hongtao

Cell reports

2022 Volume 38, Issue 7, Page(s) 110395

Abstract: Aneuploidy, defective differentiation, and inactivation of the tumor suppressor TP53 all occur frequently during tumorigenesis. Here, we probe the potential links among these cancer traits by inactivating TP53 in human embryonic stem cells (hESCs). ... ...

Abstract	Aneuploidy, defective differentiation, and inactivation of the tumor suppressor TP53 all occur frequently during tumorigenesis. Here, we probe the potential links among these cancer traits by inactivating TP53 in human embryonic stem cells (hESCs). TP53
MeSH term(s)	Amino Acid Motifs ; Animals ; CRISPR-Cas Systems/genetics ; Cell Differentiation ; Cell Line ; Cell Lineage ; Cilia/metabolism ; Cytoskeletal Proteins/genetics ; Cytoskeletal Proteins/metabolism ; Genome, Human ; Hedgehog Proteins/metabolism ; Human Embryonic Stem Cells/cytology ; Human Embryonic Stem Cells/metabolism ; Humans ; Mice, Inbred NOD ; Mice, SCID ; Neural Stem Cells/metabolism ; Neurogenesis/genetics ; Organogenesis ; Signal Transduction ; Teratoma/pathology ; Tumor Suppressor Protein p53/chemistry ; Tumor Suppressor Protein p53/metabolism ; Mice
Chemical Substances	BBS9 protein, human ; Cytoskeletal Proteins ; Hedgehog Proteins ; SHH protein, human ; Tumor Suppressor Protein p53
Language	English
Publishing date	2022-02-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2022.110395
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Article ; Online: Live-cell fluorescence imaging for phenotypic analysis of mitosis.

Sivakumar, Sushama / Daum, John R / Gorbsky, Gary J

Methods in molecular biology (Clifton, N.J.)

2014 Volume 1170, Page(s) 549–562

Abstract: Live-cell fluorescence microscopy is a powerful tool for characterizing aberrant mitotic phenotypes resulting from exposure to chemical inhibitors or after depletion of protein targets by RNA interference or other methods. Live imaging of cultured cells ... ...

Abstract	Live-cell fluorescence microscopy is a powerful tool for characterizing aberrant mitotic phenotypes resulting from exposure to chemical inhibitors or after depletion of protein targets by RNA interference or other methods. Live imaging of cultured cells during mitotic progression presents challenges in maintaining optimal health of cells while achieving the temporal and spatial resolution to accomplish the goals of the study. We describe herein strategies to monitor and analyze mammalian cell mitosis with standard, inverted, fluorescence microscopy systems that are widely available.
MeSH term(s)	Cell Culture Techniques/methods ; Cell Survival ; Green Fluorescent Proteins/analysis ; HeLa Cells ; Histones/analysis ; Humans ; Microscopy, Fluorescence/methods ; Mitosis ; Optical Imaging/methods ; Recombinant Fusion Proteins/analysis
Chemical Substances	Histones ; Recombinant Fusion Proteins ; Green Fluorescent Proteins (147336-22-9)
Language	English
Publishing date	2014-06-06
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-0888-2_31
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Inter-library loan at ZB MED

Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

Article ; Online: Ska3 Phosphorylated by Cdk1 Binds Ndc80 and Recruits Ska to Kinetochores to Promote Mitotic Progression.

Zhang, Qian / Sivakumar, Sushama / Chen, Yujue / Gao, Haishan / Yang, Lu / Yuan, Zhu / Yu, Hongtao / Liu, Hong

Current biology : CB

2017 Volume 27, Issue 10, Page(s) 1477–1484.e4

Abstract: The spindle and kinetochore-associated (Ska) protein complex is required for accurate chromosome segregation during mitosis [1-6] and consists of two copies each of Ska1, Ska2, and Ska3 proteins [4, 7]. The Ska complex contains multiple microtubule- ... ...

Abstract	The spindle and kinetochore-associated (Ska) protein complex is required for accurate chromosome segregation during mitosis [1-6] and consists of two copies each of Ska1, Ska2, and Ska3 proteins [4, 7]. The Ska complex contains multiple microtubule-binding elements and promotes kinetochore-microtubule attachment [8-11]. The Ska1 C-terminal domain (CTD) recruits protein phosphatase 1 (PP1) to kinetochores to promote timely anaphase onset [12]. The Ska complex regulates, and is regulated by, Aurora B [13]. Aurora B phosphorylates both Ska1 and Ska3 to inhibit the kinetochore localization of the Ska complex [14]. Despite its multitude of functions at kinetochores, how the Ska complex itself is recruited to kinetochores is unclear. It is unknown whether any mitotic kinases positively regulate the localization of the Ska complex to kinetochores. Here, we show that Cdk1 phosphorylates Ska3 to promote its direct binding to the Ndc80 complex (Ndc80C), a core outer kinetochore component. We also show that this phosphorylation occurs specifically during mitosis and is required for the kinetochore localization of the Ska complex. Ska3 mutants deficient in Cdk1 phosphorylation are defective in kinetochore localization but retain microtubule localization. These mutants support chromosome alignment but delay anaphase onset. We propose that Ska3 phosphorylated by Cdk1 in mitosis binds to Ndc80C and recruits the Ska complex to kinetochores where Ska1 can bind both PP1 and microtubules to promote anaphase onset.
MeSH term(s)	Aurora Kinase B/metabolism ; CDC2 Protein Kinase/metabolism ; Cell Cycle Proteins ; Chromosomal Proteins, Non-Histone/metabolism ; Chromosome Segregation ; Cytoskeletal Proteins ; HeLa Cells ; Humans ; Kinetochores/metabolism ; Microtubule-Associated Proteins/metabolism ; Microtubules/metabolism ; Mitosis ; Nuclear Proteins/metabolism ; Phosphorylation
Chemical Substances	Cell Cycle Proteins ; Chromosomal Proteins, Non-Histone ; Cytoskeletal Proteins ; Microtubule-Associated Proteins ; NDC80 protein, human ; Nuclear Proteins ; SKA1 protein, human ; SKA2 protein, human ; Ska3 protein, human ; AURKB protein, human (EC 2.7.11.1) ; Aurora Kinase B (EC 2.7.11.1) ; CDC2 Protein Kinase (EC 2.7.11.22) ; CDK1 protein, human (EC 2.7.11.22)
Language	English
Publishing date	2017-05-04
Publishing country	England
Document type	Journal Article
ZDB-ID	1071731-6
ISSN	1879-0445 ; 0960-9822
ISSN (online)	1879-0445
ISSN	0960-9822
DOI	10.1016/j.cub.2017.03.060
Database	MEDical Literature Analysis and Retrieval System OnLINE

Full text online

Accessible to users with ZB MED library card

In stock of ZB MED Cologne/Königswinter

Zs.A 3208: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (2.OG)
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾

Article ; Online: The spindle and kinetochore-associated (Ska) complex enhances binding of the anaphase-promoting complex/cyclosome (APC/C) to chromosomes and promotes mitotic exit.

Sivakumar, Sushama / Daum, John R / Tipton, Aaron R / Rankin, Susannah / Gorbsky, Gary J

Molecular biology of the cell

2014 Volume 25, Issue 5, Page(s) 594–605

Abstract: The spindle and kinetochore-associated (Ska) protein complex is a heterotrimeric complex required for timely anaphase onset. The major phenotypes seen after small interfering RNA-mediated depletion of Ska are transient alignment defects followed by ... ...

Abstract	The spindle and kinetochore-associated (Ska) protein complex is a heterotrimeric complex required for timely anaphase onset. The major phenotypes seen after small interfering RNA-mediated depletion of Ska are transient alignment defects followed by metaphase arrest that ultimately results in cohesion fatigue. We find that cells depleted of Ska3 arrest at metaphase with only partial degradation of cyclin B1 and securin. In cells arrested with microtubule drugs, Ska3-depleted cells exhibit slower mitotic exit when the spindle checkpoint is silenced by inhibition of the checkpoint kinase, Mps1, or when cells are forced to exit mitosis downstream of checkpoint silencing by inactivation of Cdk1. These results suggest that in addition to a role in fostering kinetochore-microtubule attachment and chromosome alignment, the Ska complex has functions in promoting anaphase onset. We find that both Ska3 and microtubules promote chromosome association of the anaphase-promoting complex/cyclosome (APC/C). Chromosome-bound APC/C shows significantly stronger ubiquitylation activity than cytoplasmic APC/C. Forced localization of Ska complex to kinetochores, independent of microtubules, results in enhanced accumulation of APC/C on chromosomes and accelerated cyclin B1 degradation during induced mitotic exit. We propose that a Ska-microtubule-kinetochore association promotes APC/C localization to chromosomes, thereby enhancing anaphase onset and mitotic exit.
MeSH term(s)	Anaphase/drug effects ; Anaphase/genetics ; Anaphase/physiology ; Anaphase-Promoting Complex-Cyclosome/metabolism ; Anaphase-Promoting Complex-Cyclosome/physiology ; Cell Cycle Checkpoints/drug effects ; Cell Cycle Proteins ; Chromosomes, Human/drug effects ; Chromosomes, Human/metabolism ; Cyclin B1/metabolism ; HeLa Cells ; Humans ; Metaphase/drug effects ; Metaphase/genetics ; Metaphase/physiology ; Microtubule-Associated Proteins/genetics ; Microtubule-Associated Proteins/metabolism ; Microtubule-Associated Proteins/physiology ; Mitosis/drug effects ; Mitosis/genetics ; Mitosis/physiology ; Models, Biological ; Nocodazole/pharmacology ; Tubulin Modulators/pharmacology
Chemical Substances	CCNB1 protein, human ; Cell Cycle Proteins ; Cyclin B1 ; Microtubule-Associated Proteins ; Ska3 protein, human ; Tubulin Modulators ; Anaphase-Promoting Complex-Cyclosome (EC 2.3.2.27) ; Nocodazole (SH1WY3R615)
Language	English
Publishing date	2014-01-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.E13-07-0421
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Zs.A 2853: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 410: Show issues

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article ; Online: PUMILIO hyperactivity drives premature aging of

Kopp, Florian / Elguindy, Mahmoud M / Yalvac, Mehmet E / Zhang, He / Chen, Beibei / Gillett, Frank A / Lee, Sungyul / Sivakumar, Sushama / Yu, Hongtao / Xie, Yang / Mishra, Prashant / Sahenk, Zarife / Mendell, Joshua T

eLife

2019 Volume 8

Abstract: Although numerous long noncoding RNAs (lncRNAs) have been identified, our understanding of their roles in mammalian physiology remains limited. Here, we investigated the physiologic function of the conserved ... ...

Abstract	Although numerous long noncoding RNAs (lncRNAs) have been identified, our understanding of their roles in mammalian physiology remains limited. Here, we investigated the physiologic function of the conserved lncRNA
MeSH term(s)	Aging/genetics ; Aging/physiology ; Aging, Premature/genetics ; Aging, Premature/pathology ; Animals ; Gene Expression Regulation/genetics ; Homeostasis/genetics ; Humans ; Mice ; Mitochondria/genetics ; Mitosis/genetics ; Phenotype ; RNA, Long Noncoding/genetics ; RNA-Binding Proteins/genetics ; Transcription Factors/genetics
Chemical Substances	NORAD long non-coding RNA, human ; Pum2 protein, mouse ; RNA, Long Noncoding ; RNA-Binding Proteins ; Transcription Factors
Language	English
Publishing date	2019-02-08
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.42650
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

To top

More links

Kategorien

Order via subito

Inter-library loan at ZB MED

More links

Kategorien

Order via subito

More links

Kategorien

Order via subito

More links

Kategorien

Order via subito

Inter-library loan at ZB MED

Full text online

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

Order via subito

More links

Kategorien

Order via subito

Inter-library loan at ZB MED

Full text online

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

Order via subito