title: CRISPR/Cas9-mediated deletion of lncRNA Gm26878 in the distant Foxf1 enhancer region.
abstract: Recent genome editing techniques, including CRISPR mutagenesis screens, offer unparalleled opportunities to study the regulatory non-coding genomic regions, enhancers, promoters, and functional non-coding RNAs. Heterozygous point mutations in FOXF1 and genomic deletion copy-number variants at chromosomal region 16q24.1 involving FOXF1 or its regulatory region mapping ~300 kb upstream of FOXF1 and leaving it intact have been identified in the vast majority of patients with a lethal neonatal lung disease, alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV). Homozygous Foxf1-/-mice have been shown to die by embryonic day 8.5 because of defects in the development of extraembryonic and lateral mesoderm-derived tissues, whereas heterozygous Foxf1+/-mice exhibit features resembling ACDMPV. We have previously defined a human lung-specific enhancer region encoding two long non-coding RNAs, LINC01081 and LINC01082, expressed in the lungs. To investigate the biological significance of lncRNAs in the Foxf1 enhancer region, we have generated a CRISPR/Cas9-mediated ~2.4 kb deletion involving the entire lncRNA-encoding gene Gm26878, located in the mouse region syntenic with the human Foxf1 upstream enhancer. Very recently, this mouse genomic region has been shown to function as a Foxf1 enhancer. Our results indicate that homozygous loss of Gm26878 is neonatal lethal with low penetrance. No changes in Foxf1 expression were observed, suggesting that the regulation of Foxf1 expression differs between mouse and human.
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Szafranski
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA.|Department of Genetics and Pharmaceutical Microbiology, Poznan University of Medical Sciences, Poznan, Poland.|Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.
authors.forename: Justyna A
authors.initials: JA
authors.lastname: Karolak
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Denise
authors.initials: D
authors.lastname: Lanza
authors.affiliation: Department of Genetics and Pharmaceutical Microbiology, Poznan University of Medical Sciences, Poznan, Poland.|Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.
authors.forename: Marzena
authors.initials: M
authors.lastname: Gajęcka
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA. heaney@bcm.edu.
authors.forename: Jason
authors.initials: J
authors.lastname: Heaney
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA. pawels@bcm.edu.
authors.forename: Paweł
authors.initials: P
authors.lastname: Stankiewicz
mesh_terms: D000483:Alleles; D000818:Animals; D001665:Binding Sites; D064113:CRISPR-Cas Systems; D004742:Enhancer Elements, Genetic; D051858:Forkhead Transcription Factors / Q000235:genetics*; D018628:Gene Dosage; D000072669:Gene Editing; D015870:Gene Expression; D055786:Gene Knockout Techniques; D005838:Genotype; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D016296:Mutagenesis; D010547:Persistent Fetal Circulation Syndrome / Q000235:genetics; D011485:Protein Binding; D062085:RNA, Long Noncoding / Q000235:genetics*; D017384:Sequence Deletion
chemical_list: D051858:Forkhead Transcription Factors; C486293:Foxf1 protein, mouse; D062085:RNA, Long Noncoding
keywords: Deletion Allele; Genome Editing; Transcription Factor Binding Site; UCSC Genome Browser; lncRNA Gene