Cleft palate and minor metabolic disturbances in a mouse global Arl15 gene knockout
ARL15, a small GTPase protein, was linked to metabolic traits in association studies. We aimed to test the Arl15 gene as a functional candidate for metabolic traits in the mouse. CRISPR/Cas9 germline knockout (KO) of Arl15 showed that homozygotes were postnatal lethal and exhibited a complete cleft...
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Published in | The FASEB journal Vol. 37; no. 11; pp. e23211 - n/a |
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Main Authors | , , , , , , , , , , , , , , , , , |
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Language | English |
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01.11.2023
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Abstract | ARL15, a small GTPase protein, was linked to metabolic traits in association studies. We aimed to test the Arl15 gene as a functional candidate for metabolic traits in the mouse. CRISPR/Cas9 germline knockout (KO) of Arl15 showed that homozygotes were postnatal lethal and exhibited a complete cleft palate (CP). Also, decreased cell migration was observed from Arl15 KO mouse embryonic fibroblasts (MEFs). Metabolic phenotyping of heterozygotes showed that females had reduced fat mass on a chow diet from 14 weeks of age. Mild body composition phenotypes were also observed in heterozygous mice on a high‐fat diet (HFD)/low‐fat diet (LFD). Females on a HFD showed reduced body weight, gonadal fat depot weight and brown adipose tissue (BAT) weight. In contrast, in the LFD group, females showed increased bone mineral density (BMD), while males showed a trend toward reduced BMD. Clinical biochemistry analysis of plasma on HFD showed transient lower adiponectin at 20 weeks of age in females. Urinary and plasma Mg2+ concentrations were not significantly different. Our phenotyping data showed that Arl15 is essential for craniofacial development. Adult metabolic phenotyping revealed potential roles in brown adipose tissue and bone development.
The ADP‐ribosylation factor like 15 (Arl15) gene was knocked out by using CRISPR‐Cas9 that targeted exon 2 globally. Homozygous mutant embryos exhibited a cleft palate and died postnatally. KO MEFs collected at E13.5 showed reduced cell migration in vitro. Heterozygotes were viable and exhibited minor metabolic phenotypes such as reduced fat mass on normal chow. HFD induced whitening of brown adipose tissue; however, heterozygous female mice showed less whitening compared to wildtype. LFD resulted in female heterozygous having an increased BMD. |
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AbstractList | ARL15, a small GTPase protein, was linked to metabolic traits in association studies. We aimed to test the Arl15 gene as a functional candidate for metabolic traits in the mouse. CRISPR/Cas9 germline knockout (KO) of Arl15 showed that homozygotes were postnatal lethal and exhibited a complete cleft palate (CP). Also, decreased cell migration was observed from Arl15 KO mouse embryonic fibroblasts (MEFs). Metabolic phenotyping of heterozygotes showed that females had reduced fat mass on a chow diet from 14 weeks of age. Mild body composition phenotypes were also observed in heterozygous mice on a high-fat diet (HFD)/low-fat diet (LFD). Females on a HFD showed reduced body weight, gonadal fat depot weight and brown adipose tissue (BAT) weight. In contrast, in the LFD group, females showed increased bone mineral density (BMD), while males showed a trend toward reduced BMD. Clinical biochemistry analysis of plasma on HFD showed transient lower adiponectin at 20 weeks of age in females. Urinary and plasma Mg
concentrations were not significantly different. Our phenotyping data showed that Arl15 is essential for craniofacial development. Adult metabolic phenotyping revealed potential roles in brown adipose tissue and bone development. ARL15, a small GTPase protein, was linked to metabolic traits in association studies. We aimed to test the Arl15 gene as a functional candidate for metabolic traits in the mouse. CRISPR/Cas9 germline knockout (KO) of Arl15 showed that homozygotes were postnatal lethal and exhibited a complete cleft palate (CP). Also, decreased cell migration was observed from Arl15 KO mouse embryonic fibroblasts (MEFs). Metabolic phenotyping of heterozygotes showed that females had reduced fat mass on a chow diet from 14 weeks of age. Mild body composition phenotypes were also observed in heterozygous mice on a high‐fat diet (HFD)/low‐fat diet (LFD). Females on a HFD showed reduced body weight, gonadal fat depot weight and brown adipose tissue (BAT) weight. In contrast, in the LFD group, females showed increased bone mineral density (BMD), while males showed a trend toward reduced BMD. Clinical biochemistry analysis of plasma on HFD showed transient lower adiponectin at 20 weeks of age in females. Urinary and plasma Mg2+ concentrations were not significantly different. Our phenotyping data showed that Arl15 is essential for craniofacial development. Adult metabolic phenotyping revealed potential roles in brown adipose tissue and bone development. The ADP‐ribosylation factor like 15 (Arl15) gene was knocked out by using CRISPR‐Cas9 that targeted exon 2 globally. Homozygous mutant embryos exhibited a cleft palate and died postnatally. KO MEFs collected at E13.5 showed reduced cell migration in vitro. Heterozygotes were viable and exhibited minor metabolic phenotypes such as reduced fat mass on normal chow. HFD induced whitening of brown adipose tissue; however, heterozygous female mice showed less whitening compared to wildtype. LFD resulted in female heterozygous having an increased BMD. |
Author | Bai, Ying Baaij, Jeroen H. F. Cleak, James Horner, Neil Naveenan, Navaratnam Schulze, Ulrike Pandey, Rajesh Simon, Michelle M. Bentley, Liz Mianné, Joffrey Westerberg, Henrik Ma, Chao Hough, Tertius Paphiti, Keanu Teboul, Lydia Wu, Yixing Cañas, Ramón Casero Cox, Roger D. |
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Keywords | bone mineral density adiposity intermediate filament small GTPases cell migration epithelium to mesenchymal transition (EMT) cleft palate |
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Notes | Ying Bai and Liz Bentley contributed equally to this work. |
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Snippet | ARL15, a small GTPase protein, was linked to metabolic traits in association studies. We aimed to test the Arl15 gene as a functional candidate for metabolic... |
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SubjectTerms | Adiponectin - metabolism Adipose Tissue, Brown - metabolism adiposity Animals bone mineral density cell migration cleft palate Cleft Palate - genetics Cleft Palate - metabolism Diet, High-Fat epithelium to mesenchymal transition (EMT) Female Fibroblasts - metabolism Gene Knockout Techniques intermediate filament Male Mice Mice, Inbred C57BL Mice, Knockout small GTPases |
Title | Cleft palate and minor metabolic disturbances in a mouse global Arl15 gene knockout |
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