title: Genomic safe harbors permit high β-globin transgene expression in thalassemia induced pluripotent stem cells.
abstract: Realizing the therapeutic potential of human induced pluripotent stem (iPS) cells will require robust, precise and safe strategies for genetic modification, as cell therapies that rely on randomly integrated transgenes pose oncogenic risks. Here we describe a strategy to genetically modify human iPS cells at 'safe harbor' sites in the genome, which fulfill five criteria based on their position relative to contiguous coding genes, microRNAs and ultraconserved regions. We demonstrate that ∼10% of integrations of a lentivirally encoded β-globin transgene in β-thalassemia-patient iPS cell clones meet our safe harbor criteria and permit high-level β-globin expression upon erythroid differentiation without perturbation of neighboring gene expression. This approach, combining bioinformatics and functional analyses, should be broadly applicable to introducing therapeutic or suicide genes into patient-specific iPS cells for use in cell therapy.
authors.affiliation: Center for Cell Engineering, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.
authors.forename: Eirini P
authors.initials: EP
authors.lastname: Papapetrou
authors.forename: Gabsang
authors.initials: G
authors.lastname: Lee
authors.forename: Nirav
authors.initials: N
authors.lastname: Malani
authors.forename: Manu
authors.initials: M
authors.lastname: Setty
authors.forename: Isabelle
authors.initials: I
authors.lastname: Riviere
authors.forename: Laxmi M S
authors.initials: LM
authors.lastname: Tirunagari
authors.forename: Kyuichi
authors.initials: K
authors.lastname: Kadota
authors.forename: Shoshannah L
authors.initials: SL
authors.lastname: Roth
authors.forename: Patricia
authors.initials: P
authors.lastname: Giardina
authors.forename: Agnes
authors.initials: A
authors.lastname: Viale
authors.forename: Christina
authors.initials: C
authors.lastname: Leslie
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
authors.forename: Lorenz
authors.initials: L
authors.lastname: Studer
authors.forename: Michel
authors.initials: M
authors.lastname: Sadelain
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D041905:Erythroid Cells / Q000166:cytology; D005260:Female; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D015894:Genome, Human; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D019076:Transgenes / Q000235:genetics* / Q000502:physiology; D055544:beta-Globins / Q000235:genetics / Q000378:metabolism*; D017086:beta-Thalassemia / Q000235:genetics / Q000378:metabolism / Q000628:therapy*
chemical_list: D055544:beta-Globins