TY - JOUR
T1 - An RNAi screen identifies msi2 and prox1 as having opposite roles in the regulation of hematopoietic stem cell activity
AU - Hope, Kristin
AU - Cellot, Sonia
AU - Ting, Stephen Bek Ngie
AU - MacRae, Tara
AU - Mayotte, Nadine
AU - Iscove, Norman N
AU - Sauvageau, Guy
PY - 2010
Y1 - 2010
N2 - In this study, we describe an in vivo RNA interference functional genetics approach to evaluate the role of 20 different conserved polarity factors and fate determinants in mouse hematopoietic stem cell (HSC) activity. In total, this screen revealed three enhancers and one suppressor of HSC-derived reconstitution. Pard6a, Prkcz, and Msi2 shRNAmediated depletion significantly impaired HSC repopulation. An in vitro promotion of differentiation was observed after the silencing of these genes, consistent with their function in regulating HSC selfrenewal. Conversely, Prox1 knockdown led to in vivo accumulation of primitive and differentiated cells. HSC activity was also enhanced in vitro when Prox1 levels were experimentally reduced, identifying it as a potential antagonist of self-renewal. HSC engineered to overexpress Msi2 or Prox1 showed the reverse phenotype to those transduced with corresponding shRNA vectors. Gene expression profiling studies identified a number of known HSC and cell cycle regulators as potential downstream targets to Msi2 and Prox1. A? 2010 Elsevier Inc.
AB - In this study, we describe an in vivo RNA interference functional genetics approach to evaluate the role of 20 different conserved polarity factors and fate determinants in mouse hematopoietic stem cell (HSC) activity. In total, this screen revealed three enhancers and one suppressor of HSC-derived reconstitution. Pard6a, Prkcz, and Msi2 shRNAmediated depletion significantly impaired HSC repopulation. An in vitro promotion of differentiation was observed after the silencing of these genes, consistent with their function in regulating HSC selfrenewal. Conversely, Prox1 knockdown led to in vivo accumulation of primitive and differentiated cells. HSC activity was also enhanced in vitro when Prox1 levels were experimentally reduced, identifying it as a potential antagonist of self-renewal. HSC engineered to overexpress Msi2 or Prox1 showed the reverse phenotype to those transduced with corresponding shRNA vectors. Gene expression profiling studies identified a number of known HSC and cell cycle regulators as potential downstream targets to Msi2 and Prox1. A? 2010 Elsevier Inc.
UR - http://www.sciencedirect.com.ezproxy.lib.monash.edu.au/science/article/pii/S1934590910002894
U2 - 10.1016/j.stem.2010.06.007
DO - 10.1016/j.stem.2010.06.007
M3 - Article
SN - 1934-5909
VL - 7
SP - 101
EP - 113
JO - Cell Stem Cell
JF - Cell Stem Cell
IS - 1
ER -