SARs do not impair position-dependent expression of a kit/lacZ transgene.
We have cloned and sequenced 4.2 kb of the murine c-kit promoter region. Analysis of transgenic embryos for a kit/lacZ fusion gene showed (i) an unexpected expression in the neural retina, and (ii) an unusual sensitivity to position effects. This indicates that major c-kit tissue specific regulating elements lie outside the proximal promoter. We took advantage to the intrinsic properties of the kit/lacZ transgene to investigated the potential of scaffold attachment regions (SARs) to confer position-independence on gene regulation in an in vivo context. No SAR-dependent isolation of the kit/lacZ transgene was found in the transgenic embryos, indicating a limited capacity of SAR sequences to buffer the effect of flanking sequences on transgene expression.
De Sepulveda
P
P
URA-INRA de Génétique Moleculaire, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France.
Salaün
P
P
Maas
N
N
André
C
C
Panthier
J J
JJ
D000818:Animals; D004626:Embryology; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D016899:Interferon-beta / Q000235:genetics*; D007763:Lac Operon / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics*; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics*; D016184:Receptors, Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D012160:Retina / Q000196:embryology; D014018:Tissue Distribution
D011518:Proto-Oncogene Proteins; D016184:Receptors, Colony-Stimulating Factor; D011993:Recombinant Fusion Proteins; D016899:Interferon-beta; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases