Inducible gene and shRNA expression in resident hematopoietic stem cells in vivo

Stem Cells
Elisa LaurentiAndreas Trumpp

Abstract

Hematopoietic stem cells (HSC) are probably the best understood somatic stem cells and often serve as a paradigm for other stem cells. Nevertheless, most current techniques to genetically manipulate them in vivo are either constitutive and/or induced in settings of hematopoietic stress such as after irradiation. Here, we present a conditional expression system that allows for externally controllable transgenesis and knockdown in resident HSCs, based on a lentiviral vector containing a tet-O sequence and a transgenic mouse line expressing a doxycyclin-regulated tTR-KRAB repressor protein. HSCs harvested from tTR-KRAB mice are transduced with the lentiviral vector containing a cDNA (i.e., Green Fluorescent Protein (GFP)) and/or shRNA (i.e., p53) of interest and then transplanted into lethally irradiated recipients. While the vector is effectively repressed by tTR-KRAB during homing and engraftment, robust GFP/shp53 expression is induced on doxycyclin treatment in HSCs and their progeny. Doxycylin-controllable transcription is maintained on serial transplantation, indicating that repopulating HSCs are stably modified by this approach. In summary, this easy to implement conditional system provides inducible and reversible overexpre...Continue Reading

References

Apr 1, 1995·Molecular and Cellular Biology·U DeuschleH J Thiesen
Aug 10, 2000·Molecular Therapy : the Journal of the American Society of Gene Therapy·H MizuguchiT Hayakawa
Sep 13, 2001·Proceedings of the National Academy of Sciences of the United States of America·A PfeiferI M Verma
Oct 2, 2004·PLoS Biology·Teresa A VeneziaMargaret A Goodell
Oct 4, 2005·Nature Genetics·Ross A DickinsScott W Lowe
Nov 9, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Isabelle BardeOlivier Danos
Dec 7, 2005·The Journal of Experimental Medicine·Emmanuelle PasseguéIrving L Weissman
Jan 25, 2006·Nature Methods·Jolanta SzulcPatrick Aebischer
Aug 25, 2006·Nature Methods·Maciej WiznerowiczDidier Trono
Sep 2, 2006·Proceedings of the National Academy of Sciences of the United States of America·Kum-Joo ShinIain D C Fraser
Apr 17, 2007·Brain Research·David J EveBruce A Citron
Jun 19, 2007·Nature Genetics·Ross A DickinsScott W Lowe
Sep 26, 2007·The Journal of Biological Chemistry·Maciej WiznerowiczDidier Trono
Aug 22, 2008·Nature·Laura SoucekGerard I Evan
Dec 2, 2008·Cell Stem Cell·Elisa LaurentiAndreas Trumpp
Feb 13, 2009·Nature·Marieke A G EssersAndreas Trumpp
Mar 13, 2009·Journal of Virology·Isabelle BardeDidier Trono
Apr 22, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Atsuko Yoshioka HigashiMotoko Yanagita

❮ Previous
Next ❯

Citations

Feb 27, 2013·Molecular Therapy : the Journal of the American Society of Gene Therapy·Mario AmendolaLuigi Naldini
Jul 19, 2013·Molecular Therapy. Nucleic Acids·Andrea MaddalenaSebastian Kügler
Nov 26, 2010·Nature Nanotechnology·Peixuan Guo
May 19, 2012·The Journal of Biological Chemistry·Anna C GronerDidier Trono
Sep 4, 2013·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Meikang QiuPeixuan Guo
Jul 26, 2011·Journal of Visualized Experiments : JoVE·Carmen G PaliiMarjorie Brand
Nov 26, 2013·Advanced Drug Delivery Reviews·Yi ShuPeixuan Guo
Dec 3, 2014·Journal of Molecular and Cellular Cardiology·Xinyu WengYong Xu
May 25, 2011·ACS Nano·Girish C ShuklaBruce A Shapiro
Jun 29, 2011·Biochemical and Biophysical Research Communications·Mo Sun KwonTeoan Kim
Jan 4, 2017·ACS Nano·Daniel JasinskiPeixuan Guo

❮ Previous
Next ❯

Related Concepts

Related Feeds

Blood And Marrow Transplantation

The use of hematopoietic stem cell transplantation or blood and marrow transplantation (bmt) is on the increase worldwide. BMT is used to replace damaged or destroyed bone marrow with healthy bone marrow stem cells. Here is the latest research on bone and marrow transplantation.

Adult Stem Cells

Adult stem cells reside in unique niches that provide vital cues for their survival, self-renewal, and differentiation. They hold great promise for use in tissue repair and regeneration as a novel therapeutic strategies. Here is the latest research.

Allogenic & Autologous Therapies

Allogenic therapies are generated in large batches from unrelated donor tissues such as bone marrow. In contrast, autologous therapies are manufactures as a single lot from the patient being treated. Here is the latest research on allogenic and autologous therapies.