{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,13]],"date-time":"2025-05-13T15:22:10Z","timestamp":1747149730933,"version":"3.40.5"},"reference-count":41,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T00:00:00Z","timestamp":1672876800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["JIMD Reports"],"published-print":{"date-parts":[[2023,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Asparagine synthetase (ASNS) catalyzes the synthesis of asparagine (Asn) from aspartate and glutamine. Biallelic mutations in the <jats:italic>ASNS<\/jats:italic> gene result in ASNS Deficiency (ASNSD). Children with ASNSD exhibit congenital microcephaly, epileptic\u2010like seizures, and continued brain atrophy, often leading to premature mortality. This report describes a 4\u2010year\u2010old male with global developmental delay and seizures with two novel mutations in the <jats:italic>ASNS<\/jats:italic> gene, c.614A\u2009&gt;\u2009C (maternal) and c.1192dupT (paternal) encoding p.H205P and p.Y398Lfs*4 variants, respectively. We employed the novel use of immortalized lymphoblastoid cell lines (LCL) to show that the proliferation of the heterozygotic parental LCL was not severely affected by culture in Asn\u2010free medium, but growth of the child's cells was suppressed by about 50%. Asn production by the LCL from both the father and the child was significantly decreased relative to the mother's cells. mRNA and protein analysis of the paternal LCL cells for the Y398Lfs*4 variant revealed reductions in both. Attempts to ectopically express the truncated Y398Lfs*4 variant in either HEK293T or ASNS\u2010null cells resulted in little or no detectable protein. Expression and purification of the H205P variant from HEK293T cells revealed enzymatic activity similar to wild\u2010type ASNS. Stable expression of WT ASNS rescued the growth of ASNS\u2010null JRS cells in Asn\u2010free medium and the H205P variant was only slightly less effective. However, the Y398Lfs*4 variant appeared to be unstable in JRS cells. These results indicate that co\u2010expression of the H205P and Y398Lfs*4 variants leads to a significant reduction in Asn synthesis and cellular growth.<\/jats:p>","DOI":"10.1002\/jmd2.12356","type":"journal-article","created":{"date-parts":[[2023,1,6]],"date-time":"2023-01-06T08:41:22Z","timestamp":1672994482000},"page":"167-179","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Characterizing asparagine synthetase deficiency variants in lymphoblastoid cell lines"],"prefix":"10.1002","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1746-6320","authenticated-orcid":false,"given":"Stephen J.","family":"Staklinski","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Florida College of Medicine  Gainesville Florida USA"},{"name":"School of Biological Sciences Cold Spring Harbor Laboratory, Cold Spring Harbor  New York New York 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