Table 2.
Methods for targeted gene modifications in human pluripotent stem cells
| Method | References | Cell type(s) and cell line(s) used | Targeted gene(s) | Purpose of research; comments |
|---|---|---|---|---|
| Common Homologous Recombination (HR) | Zwaka and Thomson (2003) | hES (H1.1) | HPRT1; OCT4 | Proof-of-concept for homologous recombination in hES cells |
| Urbach et al. (2004) | hES (H13) | HPRT1 | Modelling of Lesch-Nyhan syndrome in hES cells | |
| Irion et al. (2007) | hES (HES-2) | Rosa 26 human equivalent | Definition of a site suitable for gene transfer in hES cells | |
| Davis et al. (2008) | hES (HES-3) | MIXL1 | Establishment of cell lines for lineage selection in hematopoietic differentiation; Cre-lox-mediated excision of marker genes | |
| Suzuki et al. (2008) | hES (KhES1, KhES3) | HPRT1 | Proof-of-concept of adeno-associated virus (AAV)-based gene transfer for HR in hES cells | |
| Mitsui et al. (2009) | hES (KhES1; KhES3) hiPS (243H7; 246H1) | HPRT1; NANOG | Proof of concept for efficiency of HR in hES cells using adeno-associated virus-based gene transfer vectors | |
| Ruby and Zheng (2009) | hES (HUES9) | FEZF2 | Proof-of-concept for effective HR in HUES9 cells, establishment of cell lines for lineage selection of subcortical projection neurons | |
| Xue et al. (2009) | hES (BG01) | OLIG2 | Establishment of cell lines for lineage selection in neural differentiation | |
| Khan et al. (2010) | hES (H1; BG01; BG02) hiPS (several clones) | HPRT1; HMGA1; Fibroblasts subsequently used for hiPS cell generation | Proof-of-concept of AAV-based gene transfer for HR in hES and hiPS cells; hiPS cell generation from human cells subsequent to gene targeting | |
| Song et al. (2010) | hES (HUES3, HUES8, HUES9) | P53, ATM1; HPRT1 | Establishment of p53−/− and ATM−/− hES cell lines; use of bacterial artificial chromosome (BAC)-based targeting vectors | |
| Zinc finger nuclease (ZFN)—enhanced homologous recombination | Lombardo et al. (2007) | hES (HUES1, HUES3) | CCR5 | Proof-of-concept of zinc-finger nuclease (ZFN)–enhanced HR in multiple human cell types including hES cells; use of an integrase-defective lentiviral vector for gene transfer |
| Hockemeyer et al. (2009) | hES (BG01); hiPS (several clones) | Oct4; PP1R12C (AAVS1 locus); PITX3 | Proof-of-concept for zinc-finger nuclease (ZFN)–enhanced HR into expressed and non-expressed sites in hES cells | |
| Zou et al. (2009) | hES (H9, H1) hiPS (two lines) | Lentivirally transduced mutated eGFP gene; PIG-A | Proof-of-concept for zinc-finger nuclease (ZFN)–enhanced HR in hES cells; targeting of a disease-related gene in hES cells | |
| Site-specific (phage-derived) recombination systems | Di Dominico et al. (2008) | hES (H1) | HPRT1 | Establishment of a Cre/Lox P-based gene replacement system; proof-of-concept for effective recombinase-mediated cassette exchange |
| Thyagarajan et al. (2008) | hES (BG01v) | Pseudo attP sites in the human genome | Proof-of-concept for functionality of the Streptomyces phage phiC31-based recombination system in hES cells | |
| Sakurai et al. (2010) | hES (KhES1) | HPRT1 | Establishment of a Cre/Lox P-based gene replacement system |