Journal Article

CRISPR/Cas9-mediated deletion of lncRNA Gm26878 in the distant Foxf1 enhancer region

Szafranski, Przemyslaw, Gajęcka, Marzena, Heaney, Jason, Lanza, Denise, Stankiewicz, Paweł, Karolak, Justyna A.

2017

Overview

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.

• Publisher: Springer US,Springer Nature B.V
• Subject: Alleles / Alveoli / Animal Genetics and Genomics / Animals / Binding Sites / Biomedical and Life Sciences / Cell Biology / Chromosome deletion / CRISPR / CRISPR-Cas Systems / Dysplasia / Embryogenesis / Enhancer Elements, Genetic / Enhancers / Forkhead Transcription Factors - genetics / Gene deletion / Gene Dosage / Gene Editing / Gene Expression / Gene Knockout Techniques / Gene mapping / genes / Genome editing / Genotype / heterozygosity / homozygosity / Human Genetics / Humans / Kinases / Life Sciences / Lung diseases / lungs / Mesoderm / Mice / Mice, Knockout / Mutagenesis / Neonates / non-coding RNA / patients / penetrance / Persistent Fetal Circulation Syndrome - genetics / point mutation / Promoters / Protein Binding / respiratory tract diseases / RNA, Long Noncoding - genetics / Sequence Deletion / Synteny / tissues / Veins