Molecular Evidence of Genome Editing in a Mouse Model of Immunodeficiency

Scientific Reports
H H Abdul-RazakR J Yáñez-Muñoz

Abstract

Genome editing is the introduction of directed modifications in the genome, a process boosted to therapeutic levels by designer nucleases. Building on the experience of ex vivo gene therapy for severe combined immunodeficiencies, it is likely that genome editing of haematopoietic stem/progenitor cells (HSPC) for correction of inherited blood diseases will be an early clinical application. We show molecular evidence of gene correction in a mouse model of primary immunodeficiency. In vitro experiments in DNA-dependent protein kinase catalytic subunit severe combined immunodeficiency (Prkdc scid) fibroblasts using designed zinc finger nucleases (ZFN) and a repair template demonstrated molecular and functional correction of the defect. Following transplantation of ex vivo gene-edited Prkdc scid HSPC, some of the recipient animals carried the expected genomic signature of ZFN-driven gene correction. In some primary and secondary transplant recipients we detected double-positive CD4/CD8 T-cells in thymus and single-positive T-cells in blood, but no other evidence of immune reconstitution. However, the leakiness of this model is a confounding factor for the interpretation of the possible T-cell reconstitution. Our results provide supp...Continue Reading

References

Feb 10, 1983·Nature·G C BosmaM J Bosma
Oct 1, 1995·Trends in Biochemical Sciences·S P Jackson, P A Jeggo
Sep 17, 1996·Proceedings of the National Academy of Sciences of the United States of America·T BluntP A Jeggo
May 14, 1998·Gene Therapy·R J Yáñez, A C Porter
Aug 24, 1999·Gene Therapy·R J Yáñez, A C Porter
Nov 16, 2002·Nucleic Acids Research·Rafael J Yáñez, Andrew C G Porter
Jan 17, 2003·The New England Journal of Medicine·Salima Hacein-Bey-AbinaAlain Fischer
Apr 20, 2004·BioTechniques·Peter QiuGary F Gerard
Feb 24, 2006·Nature Medicine·Rafael J Yáñez-MuñozAdrian J Thrasher
Jul 3, 2007·Nature Biotechnology·Jeffrey C MillerEdward J Rebar
Oct 3, 2007·Gene Therapy·B L AlexanderS Ylä-Herttuala
Dec 17, 2008·The Journal of Clinical Investigation·Mirjam van der BurgDik C van Gent
Jun 6, 2009·Molecular Therapy : the Journal of the American Society of Gene Therapy·Klaus Wanisch, Rafael J Yáñez-Muñoz
Dec 17, 2009·Hematology·Alessandro Aiuti, Maria Grazia Roncarolo
Jan 1, 2010·The Journal of Allergy and Clinical Immunology·Luigi D Notarangelo
Jan 15, 2010·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Donald B Kohn
Aug 4, 2010·Methods in Molecular Biology·Dmitry Y GuschinEdward J Rebar
Aug 19, 2010·Nature Reviews. Genetics·Fyodor D UrnovPhilip D Gregory
Apr 26, 2011·Methods in Molecular Biology·Johan van MeerlooJacqueline Cloos
Jul 14, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Marshall W HustonGerard Wagemaker
Jun 14, 2012·Human Gene Therapy·Christine RivatAdrian J Thrasher
Sep 4, 2012·Nature Methods·Michael T CertoAndrew M Scharenberg
Jun 1, 2013·The Journal of Clinical Investigation·Lisa WoodbinePenny A Jeggo
Feb 22, 2014·Methods in Molecular Biology·Céline J RoccaRafael J Yáñez-Muñoz
Mar 7, 2014·The New England Journal of Medicine·Pablo TebasCarl H June
Apr 1, 2014·Stem Cells Translational Medicine·David A Williams, Adrian J Thrasher
May 30, 2014·Nature·Pietro GenoveseLuigi Naldini
Oct 2, 2014·The Journal of Clinical Investigation·Rajat M Gupta, Kiran Musunuru
Nov 29, 2014·Science·Jennifer A Doudna, Emmanuelle Charpentier
Aug 27, 2015·Genome Medicine·Francis RobertJerry Pelletier
Aug 9, 2016·Molecular Cell·Megan van OverbeekAndrew P May
Oct 14, 2016·Science Translational Medicine·Mark A DeWittJacob E Corn
Nov 8, 2016·Nature·Daniel P DeverMatthew H Porteus

❮ Previous
Next ❯

Citations

Oct 4, 2018·Expert Review of Clinical Immunology·Peter Olbrich, Alexandra F Freeman

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
phosphotransferase
transfection
flow cytometry
genetic modification
transfections
protein assay
electrophoresis
GeneScreen
protein kinase assay

Software Mentioned

FlowJo
FACSDiva
Image J / V2

Related Concepts

Related Feeds

CAR-T cells

Chimeric antigen receptor (CAR) T cells are cells that are genetically engineered to recognize and target specific proteins. The ability of these cells to recognize cancer antigens and eliminate tumor cells have transformed cancer immunotherapy approaches. Here is the latest research on CAR-T cells.

Related Papers

Clinical and Experimental Immunology
Alain Fischer
Advances in Experimental Medicine and Biology
Alain FischerMarina Cavazzana-Calvo
Transfusion clinique et biologique : journal de la Société française de transfusion sanguine
Marina Cavazzana-CalvoA Fischer
© 2022 Meta ULC. All rights reserved