{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T01:25:15Z","timestamp":1773797115773,"version":"3.50.1"},"reference-count":21,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,2,6]],"date-time":"2023-02-06T00:00:00Z","timestamp":1675641600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"University of Manitoba Collaborative Research Program","award":["54302"],"award-info":[{"award-number":["54302"]}]},{"name":"University of Manitoba Collaborative Research Program","award":["RCN-160422"],"award-info":[{"award-number":["RCN-160422"]}]},{"name":"University of Manitoba Collaborative Research Program","award":["SMG2021-01"],"award-info":[{"award-number":["SMG2021-01"]}]},{"name":"Canada Institute of Health Research (CIHR)","award":["54302"],"award-info":[{"award-number":["54302"]}]},{"name":"Canada Institute of Health Research (CIHR)","award":["RCN-160422"],"award-info":[{"award-number":["RCN-160422"]}]},{"name":"Canada Institute of Health Research (CIHR)","award":["SMG2021-01"],"award-info":[{"award-number":["SMG2021-01"]}]},{"name":"CHRIM Small Operating grant","award":["54302"],"award-info":[{"award-number":["54302"]}]},{"name":"CHRIM Small Operating grant","award":["RCN-160422"],"award-info":[{"award-number":["RCN-160422"]}]},{"name":"CHRIM Small Operating grant","award":["SMG2021-01"],"award-info":[{"award-number":["SMG2021-01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>PRUNE1 is a member of the aspartic acid-histidine-histidine (DHH) protein superfamily, which could display an exopolyphosphatase activity and interact with multiple cellular proteins involved in the cytoskeletal rearrangement. It is widely expressed during embryonic development and is essential for embryogenesis. PRUNE1 could also be critical for postnatal development of the nervous system as it was found to be mutated in patients with microcephaly, brain malformations, and neurodegeneration. To determine the cellular function of PRUNE1 during development and in disease, we have generated conditional mouse alleles of the Prune1 in which loxP sites flank exon 6. Crossing these alleles with a ubiquitous Cre transgenic line resulted in a complete loss of PRUNE1 expression and embryonic defects identical to those previously described for Prune1 null embryos. In addition, breeding these alleles with a Purkinje cell-specific Cre line (Pcp2-Cre) resulted in the loss of Purkinje cells similar to that observed in patients carrying a mutation with loss of PRUNE1 function. Therefore, the Prune1 conditional mouse alleles generated in this study provide important genetic tools not only for dissecting the spatial and temporal roles of PRUNE1 during development but also for understanding the pathogenic role of PRUNE1 dysfunction in neurodegenerative or neurodevelopmental disease. In addition, from this work, we have described an approach that allows one to efficiently generate conditional mouse alleles based on mouse zygote electroporation.<\/jats:p>","DOI":"10.3390\/cells12040524","type":"journal-article","created":{"date-parts":[[2023,2,6]],"date-time":"2023-02-06T04:25:28Z","timestamp":1675657528000},"page":"524","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Generation of Conditional Knockout Alleles for PRUNE-1"],"prefix":"10.3390","volume":"12","author":[{"given":"Xiaoli","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB R3E 0J9, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Louise R.","family":"Simard","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB R3E 0J9, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hao","family":"Ding","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB R3E 0J9, Canada"},{"name":"Children\u2019s Hospital Research Institute of Manitoba (CHRIM), Winnipeg, MB R3E 0Z3, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"9707","DOI":"10.1021\/bi8010847","article-title":"Human metastasis regulator protein H-prune is a short-chain exopolyphosphatase","volume":"47","author":"Tammenkoski","year":"2008","journal-title":"Biochemistry"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3516","DOI":"10.1093\/hmg\/ddaa237","article-title":"NMIHBA results from hypomorphic PRUNE1 variants that lack short-chain exopolyphosphatase activity","volume":"29","author":"Nistala","year":"2020","journal-title":"Hum. Mol. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/S1535-6108(04)00021-2","article-title":"Prune cAMP phosphodiesterase binds nm23-H1 and promotes cancer metastasis","volume":"5","author":"Garzia","year":"2004","journal-title":"Cancer Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"7244","DOI":"10.1038\/sj.onc.1203140","article-title":"Evidence for interaction between human PRUNE and nm23-H1 NDPKinase","volume":"18","author":"Reymond","year":"1999","journal-title":"Oncogene"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"898","DOI":"10.1128\/MCB.26.3.898-911.2006","article-title":"Glycogen Synthase Kinase 3 and h-prune Regulate Cell Migration by Modulating Focal Adhesions","volume":"26","author":"Kobayashi","year":"2006","journal-title":"Mol. Cell. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1093\/brain\/awx014","article-title":"PRUNE is crucial for normal brain development and mutated in microcephaly with neurodevelopmental impairment","volume":"140","author":"Zollo","year":"2017","journal-title":"Brain"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"758146","DOI":"10.3389\/fonc.2021.758146","article-title":"Functional Genomics of PRUNE1 in Neurodevelopmental Disorders (NDDs) Tied to Medulloblastoma (MB) and Other Tumors","volume":"11","author":"Sorice","year":"2021","journal-title":"Front. Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1111\/cge.13385","article-title":"PRUNE1-related disorder: Expanding the clinical spectrum","volume":"94","author":"Imagawa","year":"2018","journal-title":"Clin. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1002\/ajmg.a.60690","article-title":"A homozygous canonical splice acceptor site mutation in PRUNE1 is responsible for a rare childhood neurodegenerative disease in Manitoba Cree families","volume":"179","author":"Hartley","year":"2019","journal-title":"Am. J. Med. Genet. Part A"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1868","DOI":"10.1002\/ajmg.a.62704","article-title":"PRUNE1 c. 933G >A synonymous variant induces exon 7 skipping, disrupts the DHHA2 domain, and leads to an atypical NMIHBA syndrome presentation: Case report and review of the literature","volume":"188","author":"Magyar","year":"2022","journal-title":"Am. J. Med. Genet. Part A"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1002\/ajmg.a.38066","article-title":"Homozygous mutation in PRUNE1 in an Oji-Cree male with a complex neurological phenotype","volume":"173","author":"Costain","year":"2017","journal-title":"Am. J. Med. Genet. Part A"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ydbio.2016.07.017","article-title":"Electroporation of Cas9 protein\/sgRNA into early pronuclear zygotes generates non-mosaic mutants in the mouse","volume":"418","author":"Hashimoto","year":"2016","journal-title":"Dev. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"14457","DOI":"10.1074\/jbc.M116.733154","article-title":"Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes","volume":"291","author":"Chen","year":"2016","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.ymeth.2021.01.005","article-title":"Production of knockout mouse lines with Cas9","volume":"191","author":"Gertsenstein","year":"2021","journal-title":"Methods"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5860","DOI":"10.1073\/pnas.93.12.5860","article-title":"Efficient in vivo manipulation of mouse genomic sequences at the zygote stage","volume":"93","author":"Lakso","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1002\/gene.20062","article-title":"Highly restricted expression of Cre recombinase in cerebellar Purkinje cells","volume":"40","author":"Zhang","year":"2004","journal-title":"Genes"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Wu, X., Sandhu, S., Patel, N., Triggs-Raine, B., and Ding, H. (2010). EMG1 is essential for mouse pre-implantation embryo development. BMC Dev. Biol., 10.","DOI":"10.1186\/1471-213X-10-99"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/1476-4598-11-18","article-title":"Loss of HLTF function promotes intestinal carcinogenesis","volume":"11","author":"Sandhu","year":"2012","journal-title":"Mol. Cancer"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"D988","DOI":"10.1093\/nar\/gkab1049","article-title":"Ensembl 2022","volume":"50","author":"Cunningham","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1038\/nrm4063","article-title":"Nonsense-mediated mRNA decay: An intricate machinery that shapes transcriptomes","volume":"16","author":"Jensen","year":"2015","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1007\/s10863-006-9044-z","article-title":"PRUNE and NM23-M1 expression in embryonic and adult mouse brain","volume":"38","author":"Carotenuto","year":"2006","journal-title":"J. Bioenerg. Biomembr."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/4\/524\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:25:09Z","timestamp":1760120709000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/4\/524"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,6]]},"references-count":21,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["cells12040524"],"URL":"https:\/\/doi.org\/10.3390\/cells12040524","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,6]]}}}