title: Non-coding-regulatory regions of human brain genes delineated by bacterial artificial chromosome knock-in mice.
abstract: BACKGROUND
  The next big challenge in human genetics is understanding the 98% of the genome that comprises non-coding DNA. Hidden in this DNA are sequences critical for gene regulation, and new experimental strategies are needed to understand the functional role of gene-regulation sequences in health and disease. In this study, we build upon our HuGX ('high-throughput human genes on the X chromosome') strategy to expand our understanding of human gene regulation in vivo.
  
  
  RESULTS
  In all, ten human genes known to express in therapeutically important brain regions were chosen for study. For eight of these genes, human bacterial artificial chromosome clones were identified, retrofitted with a reporter, knocked single-copy into the Hprt locus in mouse embryonic stem cells, and mouse strains derived. Five of these human genes expressed in mouse, and all expressed in the adult brain region for which they were chosen. This defined the boundaries of the genomic DNA sufficient for brain expression, and refined our knowledge regarding the complexity of gene regulation. We also characterized for the first time the expression of human MAOA and NR2F2, two genes for which the mouse homologs have been extensively studied in the central nervous system (CNS), and AMOTL1 and NOV, for which roles in CNS have been unclear.
  
  
  CONCLUSIONS
  We have demonstrated the use of the HuGX strategy to functionally delineate non-coding-regulatory regions of therapeutically important human brain genes. Our results also show that a careful investigation, using publicly available resources and bioinformatics, can lead to accurate predictions of gene expression.
authors.affiliation: Centre for Molecular Medicine and Therapeutics at the Child and Family Research Institute, University of British Columbia, Vancouver, British Columbia V5Z 4H4, Canada. simpson@cmmt.ubc.ca.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Schmouth
authors.forename: Mauro
authors.initials: M
authors.lastname: Castellarin
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Laprise
authors.forename: Kathleen G
authors.initials: KG
authors.lastname: Banks
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Bonaguro
authors.forename: Simone C
authors.initials: SC
authors.lastname: McInerny
authors.forename: Lisa
authors.initials: L
authors.lastname: Borretta
authors.forename: Mahsa
authors.initials: M
authors.lastname: Amirabbasi
authors.forename: Andrea J
authors.initials: AJ
authors.lastname: Korecki
authors.forename: Elodie
authors.initials: E
authors.lastname: Portales-Casamar
authors.forename: Gary
authors.initials: G
authors.lastname: Wilson
authors.forename: Lisa
authors.initials: L
authors.lastname: Dreolini
authors.forename: Steven J M
authors.initials: SJ
authors.lastname: Jones
authors.forename: Wyeth W
authors.initials: WW
authors.lastname: Wasserman
authors.forename: Daniel
authors.initials: D
authors.lastname: Goldowitz
authors.forename: Robert A
authors.initials: RA
authors.lastname: Holt
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Simpson
mesh_terms: D025042:Amacrine Cells / Q000166:cytology / Q000378:metabolism; D000091082:Angiomotins; D000818:Animals; D001921:Brain / Q000378:metabolism*; D051837:COUP Transcription Factor II / Q000235:genetics; D002490:Central Nervous System / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D041321:Chromosomes, Human, X / Q000235:genetics*; D019295:Computational Biology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D017353:Gene Deletion; D005786:Gene Expression Regulation; D055879:Gene Knock-In Techniques; D056426:Genetic Loci; D015894:Genome, Human; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008995:Monoamine Oxidase / Q000235:genetics; D055514:Nephroblastoma Overexpressed Protein / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D000091082:Angiomotins; D051837:COUP Transcription Factor II; D008565:Membrane Proteins; D008995:Monoamine Oxidase; D055514:Nephroblastoma Overexpressed Protein; C510063:AMOTL1 protein, human; C526319:CCN3 protein, human; C579458:monoamine oxidase A, human; C095569:NR2F2 protein, human