Analysis of vertebrate SCL loci identifies conserved enhancers

• Published in: Nature biotechnology Vol. 18; no. 2; pp. 181 - 186
• Main Authors: Green, Anthony R, Göttgens, Berthold, Barton, Linda M, Gilbert, James G.R, Bench, Anthony J, Sanchez, Maria-José, Bahn, Sabine, Mistry, Shailesh, Grafham, Darren, McMurray, Amanda, Vaudin, Mark, Amaya, Enrique, Bentley, David R
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.02.2000; Nature Publishing Group
• Subjects: Amino Acid Sequence; Animals; Animals, Genetically Modified; Base Sequence; Basic Helix-Loop-Helix Transcription Factors; Bioassays; Biological and medical sciences; Chickens; Conserved Sequence; Deoxyribonucleic acid; DNA; DNA-Binding Proteins - genetics; Enhancer Elements, Genetic; Fundamental and applied biological sciences. Psychology; Helix-Loop-Helix Motifs; high-throughput screening; Humans; Mammals; Mice; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Poultry; Proto-Oncogene Proteins; Rhombencephalon - embryology; Sequence Homology, Amino Acid; SLC gene; T-Cell Acute Lymphocytic Leukemia Protein 1; Transcription Factors - genetics; Transcription. Transcription factor. Splicing. Rna processing; Vertebrates - genetics; Xenopus; Xenopus Proteins; Human; Nucleotide sequence; Transcription promoter; Xenopus laevis; Rodentia; Amphibia; Transgenic animal; Homology; Salientia; Gene expression; Angiogenesis; Vertebrata; Mammalia; Reporter gene; Mouse; Hematopoiesis; Development; Regulatory sequence; Chicken; Aves; Transcription factor
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/72635

The SCL gene encodes a highly conserved bHLH transcription factor with a pivotal role in hemopoiesis and vasculogenesis. We have sequenced and analyzed 320 kb of genomic DNA composing the SCL loci from human, mouse, and chicken. Long-range sequence comparisons demonstrated multiple peaks of human/mouse homology, a subset of which corresponded precisely with known SCL enhancers. Comparisons between mammalian and chicken sequences identified some, but not all, SCL enhancers. Moreover, one peak of human/mouse homology (+23 region), which did not correspond to a known enhancer, showed significant homology to an analogous region of the chicken SCL locus. A transgenic Xenopus reporter assay was established and demonstrated that the +23 region contained a new neural enhancer. This combination of long-range comparative sequence analysis with a high-throughput transgenic bioassay provides a powerful strategy for identifying and characterizing developmentally important enhancers.