Table 1.
Genetic loci successfully targeted in hESC5.
| Targeted locus* | Year | Expressed in hESC | Cell line (sex) | Homology arms 5′/3′ (Kb) | Targeting location | Positive selection† | Negative selection‡ | Reporter cell lines§ | RMCE | Recombinase |
|---|---|---|---|---|---|---|---|---|---|---|
| HPRT | 2003 8 | Yes | H1(XY) | 1.9/10 | Exons 7–9 | Neo | TK and 6‐TG | |||
| 2004 19 | Yes | NR¶(XY) | 8.5 total | Exon 2 | Hygro | 6‐TG | ||||
| 2008 22 | Yes | H1(XY) | 4.8/2.1 | Exon 3 | EF1α‐Neo | 6‐TG | yes | CAG‐Cre | ||
| 2010 24 | Yes | KhES1(XX) | 7/2 | Exon 1 | PGK‐Neo | – | yes | EF1α‐Cre | ||
| 2010 14 | Yes | HUES‐3(XY) and HUES‐8(XY) | 118/77 | Intron 6 | CAG‐Neo‐IRES‐Puro | – | ||||
| OCT4 | 2003 8 | Yes | H1.1(XY) | 6.3/1.6 and 6.3/6.5 | Exons 3–5 | Neo | – | YES (EGFP) | ||
| ROSA26 | 2007 23 | Yes | HES‐2(XX) | 1.0/4.1 | Intron 1 | Neo | DTA | YES | Cre | |
| MIXL1 | 2008 9 | No | HES‐3(XX) | 9.4/1.9 | Exon 1 | PGK‐Neo | – | YES (GFP) | Cre | |
| FEZF2 | 2009 18 | No | HUES‐9(XX) | 4.2/4.1 | Exon 2 | PGK‐Neo | TK | YES (EYFP) | Cre | |
| OL1G2 | 2009 25 | No | BG01(XY) | 2/5.3 | Exon 2 | PGK‐Neo | TK | YES (EGFP) | Cre | |
| ISL1 | 2009 16 | No | H9(XX) HUES‐3(XX) | 9.2/5.5 | Exon 1 | Puro | – | YES (dsRED) | Cre | |
| ATM | 2010 14 | Yes | HUES‐9(XX) and H9(XX) | 9.0/2.0 and 103/6.5 | Intron 40 | CAG‐Neo CAG‐Neo‐IRES‐Puro | ||||
| p53 | 2010 14 | Yes | Not stated | 77/5.5 | Exons 2–6 | Same as ATM | – | |||
| BAF25Oa | 2010 15 | Yes | H9(XX) HUES‐8 (XY) | 3.9/3.7 | Intron 8 | β‐geo | – | CAG‐FIp Cre‐ERT2 | ||
| NANOG | 2010 17 | Yes | HUES‐1(XX) HUES‐3(XY) | 12.5/3.5 | Exon 1; includes 5′‐UTR | SV40‐Neo | – | YES (EGFP) |
*HPRT = hypoxanthine polyribosyl transferase. †Promoters indicated if used; Neo = neomycin (G418); Hygro = hygromycin; CAG = cytomegalovirus early enhancer element and chicken β‐actin promoter; β‐geo =β‐galactosidase‐neomycin resistance fusion gene. ‡TK = thymidine kinase; 6‐TG = 6‐thioguanine; DTA = diphtheria toxin A. §EGFP = enhanced green fluorescent protein; EYFP = enhanced yellow fluorescent protein; dsRED = red fluorescent protein. ¶NR = not reported. RMCE = recombinase‐mediated cassette exchange.