Functional Assessment of a New PBX1 Variant in a 46,XY Fetus with Severe Syndromic Difference of Sexual Development through CRISPR-Cas9 Gene Editing

Sexual development is a complex process relying on numerous genes. Disruptions in some of these genes are known to cause differences of sexual development (DSDs). Advances in genome sequencing allowed the discovery of new genes implicated in sexual development, such as PBX1. We present here a fetus...

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Bibliographic Details
Published in:Genes 2023-01, Vol.14 (2), p.273
Main Authors: Mary, Laura, Leclerc, Delphine, Labalme, Audrey, Bellaud, Pascale, Mazaud-Guittot, Séverine, Dréano, Stéphane, Evrard, Bertrand, Bigand, Antoine, Cauchoix, Aurélie, Loget, Philippe, Lokchine, Anna, Cluzeau, Laurence, Gilot, David, Belaud-Rotureau, Marc-Antoine, Jaillard, Sylvie
Format: Article
Language:English
Subjects:
Antibodies
Cell culture
Cell lines
Cell proliferation
Chromosomes
CRISPR
CRISPR-Cas Systems
DNA sequencing
Fetus
Fetuses
Gene Editing
Genes
Genetic aspects
Genetics
Genomes
Genomics
Genotype & phenotype
HEK293 Cells
Humans
Life Sciences
Methods
Mutants
Mutation
Nucleotide sequencing
Patients
Phenotypes
Physiological aspects
Plasmids
Pre-B-Cell Leukemia Transcription Factor 1 - genetics
Proteins
Sexual Development
Software
Splicing factors
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Summary:Sexual development is a complex process relying on numerous genes. Disruptions in some of these genes are known to cause differences of sexual development (DSDs). Advances in genome sequencing allowed the discovery of new genes implicated in sexual development, such as PBX1. We present here a fetus with a new PBX1 NM_002585.3: c.320G>A,p.(Arg107Gln) variant, presenting with severe DSD along with renal and lung malformations. Using CRISPR-Cas9 gene editing on HEK293T cells, we generated a KD cell line for PBX1. The KD cell line showed reduced proliferation and adhesion properties compared with HEK293T cells. HEK293T and KD cells were then transfected plasmids coding either PBX1 WT or PBX1-320G>A (mutant). WT or mutant PBX1 overexpression rescued cell proliferation in both cell lines. RNA-seq analyses showed less than 30 differentially expressed genes, in ectopic mutant-PBX1-expressing cells compared with WT-PBX1. Among them, , encoding a splicing factor subunit, is an interesting candidate. Overall, mutant PBX1 seems to have modest effects compared with WT PBX1 in our model. However, the recurrence of PBX1 Arg107 substitution in patients with closely related phenotypes calls for its impact in human diseases. Further functional studies are needed to explore its effects on cellular metabolism.
ISSN:2073-4425
2073-4425
DOI:10.3390/genes14020273