PUBLICATION
Establishment of iPSC lines and zebrafish with loss-of-function AHDC1 variants: models for Xia-Gibbs syndrome
- Authors
- Machado Lara Carvalho, L., Varella Branco, E., Delgado Sarafian, R., Shigeru Kobayashi, G., Tófoli de Araújo, F., Santos Souza, L., de Paula Moreira, D., Shih Ping Hsia, G., Maria Goloni Bertollo, E., Barbosa Buck, C., Souza da Costa, S., Mendes Fialho, D., Tadeu Galante Rocha de Vasconcelos, F., Abreu Brito, L., Elena de Souza Fraga Machado, L., Cabreira Ramos, I., da Veiga Pereira, L., Priszkulnik Koiffmann, C., Rita Dos Santos E Passos-Bueno, M., Antonio de Oliveira Mendes, T., Cristina Victorino Krepischi, A., Rosenberg, C.
- ID
- ZDB-PUB-230414-71
- Date
- 2023
- Source
- Gene 871: 147424 (Journal)
- Registered Authors
- Machado Lara Carvalho, Laura
- Keywords
- AHDC1, CRISPR–Cas9, Xia-Gibbs syndrome, iPSC, neurodevelopment, zebrafish
- MeSH Terms
-
- Abnormalities, Multiple*/genetics
- Animals
- Cell Differentiation/genetics
- Induced Pluripotent Stem Cells*/metabolism
- Intellectual Disability*/genetics
- Leukocytes, Mononuclear
- Syndrome
- Zebrafish/genetics
- PubMed
- 37054903 Full text @ Gene
Citation
Machado Lara Carvalho, L., Varella Branco, E., Delgado Sarafian, R., Shigeru Kobayashi, G., Tófoli de Araújo, F., Santos Souza, L., de Paula Moreira, D., Shih Ping Hsia, G., Maria Goloni Bertollo, E., Barbosa Buck, C., Souza da Costa, S., Mendes Fialho, D., Tadeu Galante Rocha de Vasconcelos, F., Abreu Brito, L., Elena de Souza Fraga Machado, L., Cabreira Ramos, I., da Veiga Pereira, L., Priszkulnik Koiffmann, C., Rita Dos Santos E Passos-Bueno, M., Antonio de Oliveira Mendes, T., Cristina Victorino Krepischi, A., Rosenberg, C. (2023) Establishment of iPSC lines and zebrafish with loss-of-function AHDC1 variants: models for Xia-Gibbs syndrome. Gene. 871:147424.
Abstract
Xia-Gibbs syndrome (XGS) is a syndromic form of intellectual disability caused by heterozygous AHDC1 variants, but the pathophysiological mechanisms underlying this syndrome are still unclear. In this manuscript, we describe the development of two different functional models: three induced pluripotent stem cell (iPSC) lines with different loss-of-function (LoF) AHDC1 variants, derived by reprogramming peripheral blood mononuclear cells from XGS patients, and a zebrafish strain with a LoF variant in the ortholog gene (ahdc1) obtained through CRISPR/Cas9-mediated editing. The three iPSC lines showed expression of pluripotency factors (SOX2, SSEA-4, OCT3/4, and NANOG). To verify the capacity of iPSC to differentiate into the three germ layers, we obtained embryoid bodies (EBs), induced their differentiation, and confirmed the mRNA expression of ectodermal, mesodermal, and endodermal markers using the TaqMan hPSC Scorecard. The iPSC lines were also approved for the following quality tests: chromosomal microarray analysis (CMA), mycoplasma testing, and short tandem repeat (STR) DNA profiling. The zebrafish model has an insertion of four base pairs in the ahdc1 gene, is fertile, and breeding between heterozygous and wild-type (WT) animals generated offspring in a genotypic proportion in agreement with Mendelian law. The established iPSC and zebrafish lines were deposited on the hpscreg.eu and zfin.org platforms, respectively. These biological models are the first for XGS and will be used in future studies that investigate the pathophysiology of this syndrome, unraveling its underlying molecular mechanisms.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping