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1. An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes

Author

Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, UK10K Consortium, Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, and UK10K Consortium

Published
2015

2. An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes

Author

Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, UK10K Consortium, Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, and UK10K Consortium

Published
2015

3. An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes

Author

Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, UK10K Consortium, Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, and UK10K Consortium

Published
2015

4. An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes

Author

Regenerative Medicine and Stem Cells, Nefro Vasculaire Geneeskunde, Circulatory Health, Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, UK10K Consortium, Regenerative Medicine and Stem Cells, Nefro Vasculaire Geneeskunde, Circulatory Health, Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A, Szymanska, Katarzyna, Nguyen, Thanh-Minh T, Racher, Hilary, Phelps, Ian G, Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A, Sorusch, Nasrin, Abdelhamed, Zakia A, Natarajan, Subaashini, Herridge, Warren, van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A, Letteboer, Stef J F, Roosing, Susanne, Adams, Matthew, Bell, Sandra M, Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E, Tomlinson, Darren C, Slaats, Gisela G, van Dam, Teunis J P, Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V, Boyle, Evan A, Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A, Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A, Chodirker, Bernard N, Chudley, Albert E, Lamont, Ryan, Bernier, Francois P, Beaulieu, Chandree L, Gordon, Paul, Pon, Richard T, Donahue, Clem, Giles, Rachel H, and UK10K Consortium

Published
2015

5. () Immunoblot showing TSC1 and TSC2 in 97-1 Neo control and FLAG-tagged wild-type and mutant TSC1 cell line lysates immunoprecipitated with anti-TSC1 and non-specific mouse IgM antibodies

Author

Louis S. Pymar, Fiona M. Platt, Jon M. Askham, Ewan E. Morrison, Margaret A. Knowles, Louis S. Pymar, Fiona M. Platt, Jon M. Askham, Ewan E. Morrison, and Margaret A. Knowles

Abstract

() Immunoblot showing expression levels of GFP-tagged wild-type and missense mutant TSC1 protein in HCV29 cell lines. Also shown are levels of S6 phosphorylation in transiently amino acid starved cells.Copyright information:Taken from "Bladder tumour-derived somatic missense mutations cause loss of function via distinct mechanisms"Human Molecular Genetics 2008;17(13):2006-2017.Published online 7 Apr 2008PMCID:PMC2427143.© The Author 2008

Published
2011
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6. Human ASPM participates in spindle organisation, spindle orientation and cytokinesis

Author

Higgins, Julie, Midgley, Carol A., Bergh, Anna-Maria, Bell, Sandra M., Askham, Jonathan M., Roberts, Emma, Binns, Ruth K., Sharif, Saghira M., Bennett, Christopher, Glover, David M., Woods, C. Geoffrey, MorrisonBond, Ewan E., Bond, Jacquelyn, Higgins, Julie, Midgley, Carol A., Bergh, Anna-Maria, Bell, Sandra M., Askham, Jonathan M., Roberts, Emma, Binns, Ruth K., Sharif, Saghira M., Bennett, Christopher, Glover, David M., Woods, C. Geoffrey, MorrisonBond, Ewan E., and Bond, Jacquelyn

Abstract

Background Mutations in the Abnormal Spindle Microcephaly related gene (ASPM) are the commonest cause of autosomal recessive primary microcephaly (MCPH) a disorder characterised by a small brain and associated mental retardation. ASPM encodes a mitotic spindle pole associated protein. It is suggested that the MCPH phenotype arises from proliferation defects in neural progenitor cells (NPC). Results We show that ASPM is a microtubule minus end-associated protein that is recruited in a microtubule-dependent manner to the pericentriolar matrix (PCM) at the spindle poles during mitosis. ASPM siRNA reduces ASPM protein at the spindle poles in cultured U2OS cells and severely perturbs a number of aspects of mitosis, including the orientation of the mitotic spindle, the main determinant of developmental asymmetrical cell division. The majority of ASPM depleted mitotic cells fail to complete cytokinesis. In MCPH patient fibroblasts we show that a pathogenic ASPM splice site mutation results in the expression of a novel variant protein lacking a tripeptide motif, a minimal alteration that correlates with a dramatic decrease in ASPM spindle pole localisation. Moreover, expression of dominant-negative ASPM C-terminal fragments cause severe spindle assembly defects and cytokinesis failure in cultured cells. Conclusions These observations indicate that ASPM participates in spindle organisation, spindle positioning and cytokinesis in all dividing cells and that the extreme C-terminus of the protein is required for ASPM localisation and function. Our data supports the hypothesis that the MCPH phenotype caused by ASPM mutation is a consequence of mitotic aberrations during neurogenesis. We propose the effects of ASPM mutation are tolerated in somatic cells but have profound consequences for the symmetrical division of NPCs, due to the unusual morphology of these cells. This antagonises the early expansion of the progenitor

7. Human ASPM participates in spindle organisation, spindle orientation and cytokinesis

Author

Higgins, Julie, Midgley, Carol A., Bergh, Anna-Maria, Bell, Sandra M., Askham, Jonathan M., Roberts, Emma, Binns, Ruth K., Sharif, Saghira M., Bennett, Christopher, Glover, David M., Woods, C. Geoffrey, MorrisonBond, Ewan E., Bond, Jacquelyn, Higgins, Julie, Midgley, Carol A., Bergh, Anna-Maria, Bell, Sandra M., Askham, Jonathan M., Roberts, Emma, Binns, Ruth K., Sharif, Saghira M., Bennett, Christopher, Glover, David M., Woods, C. Geoffrey, MorrisonBond, Ewan E., and Bond, Jacquelyn

Abstract

Background Mutations in the Abnormal Spindle Microcephaly related gene (ASPM) are the commonest cause of autosomal recessive primary microcephaly (MCPH) a disorder characterised by a small brain and associated mental retardation. ASPM encodes a mitotic spindle pole associated protein. It is suggested that the MCPH phenotype arises from proliferation defects in neural progenitor cells (NPC). Results We show that ASPM is a microtubule minus end-associated protein that is recruited in a microtubule-dependent manner to the pericentriolar matrix (PCM) at the spindle poles during mitosis. ASPM siRNA reduces ASPM protein at the spindle poles in cultured U2OS cells and severely perturbs a number of aspects of mitosis, including the orientation of the mitotic spindle, the main determinant of developmental asymmetrical cell division. The majority of ASPM depleted mitotic cells fail to complete cytokinesis. In MCPH patient fibroblasts we show that a pathogenic ASPM splice site mutation results in the expression of a novel variant protein lacking a tripeptide motif, a minimal alteration that correlates with a dramatic decrease in ASPM spindle pole localisation. Moreover, expression of dominant-negative ASPM C-terminal fragments cause severe spindle assembly defects and cytokinesis failure in cultured cells. Conclusions These observations indicate that ASPM participates in spindle organisation, spindle positioning and cytokinesis in all dividing cells and that the extreme C-terminus of the protein is required for ASPM localisation and function. Our data supports the hypothesis that the MCPH phenotype caused by ASPM mutation is a consequence of mitotic aberrations during neurogenesis. We propose the effects of ASPM mutation are tolerated in somatic cells but have profound consequences for the symmetrical division of NPCs, due to the unusual morphology of these cells. This antagonises the early expansion of the progenitor

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