A newly identified gene Ahed plays essential roles in haematopoiesis through RNA splicing

A newly identified gene Ahed plays essential roles in haematopoiesis through RNA splicing | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Help Center Sign In Submit a Preprint Cite Share Download PDF Article A newly identified gene Ahed plays essential roles in haematopoiesis through RNA splicing Takafumi Yokota, Ritsuko Nakai, Masahiro Tokunaga, Mikiro Takaishi, and 15 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2431052/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 25 Jun, 2024 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract The development of haematopoiesis is regulated by coordinated actions of multiple genes. Mutations in some genes are intimately involved in the occurrence of haematological cancers. Here, we report a previously undefined functional gene in haematopoiesis, which we isolated by screening mutant embryonic stem cells. The gene, named “ attenuated haematopoietic development ” ( Ahed ), encoded an uncharacterized nuclear protein. We found that Ahed conditional knock out (cKO) foetuses became anaemic after E14.5 and died before birth. Transplantation experiments revealed that Ahed -deficient lineage - Sca-1 + c-Kit Hi cells were unable to reconstitute haematopoiesis in vivo , suggesting that Ahed is indispensable for functional haematopoietic stem/progenitor cells. Multiple RNA sequencing analyses of Ahed -cKO cells revealed that this protein governs various critical pathways by regulating alternative splicing of RNAs. Published databases of cancer-related mutations present blood cancer patients with somatic mutations in the Ahed gene. Thus, Ahed plays a fundamental role in haematopoiesis through RNA splicing, whose mutations relates to malignancies. Biological sciences/Developmental biology/Haematopoiesis/Lymphopoiesis Biological sciences/Genetics/Development Biological sciences/Stem cells/Haematopoietic stem cells Biological sciences/Genetics/RNA splicing Biological sciences/Stem cells/Embryonic stem cells Ahed haematopoiesis mutant embryonic stem cells screening nuclear protein RNA splicing C11orf57 Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryData1.xlsx Dataset 1 NakaiAhedSupplementaryInformation.pdf Cite Share Download PDF Status: Published Journal Publication published 25 Jun, 2024 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2431052","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":225606627,"identity":"dcc9445f-ed22-436e-9248-c7161ecd1e9a","order_by":0,"name":"Takafumi 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Watanabe","email":"","orcid":"","institution":"Laboratory of Animal Experiments for Regeneration, Institute for Frontier Medical Sciences, Kyoto University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hitomi","middleName":"","lastName":"Watanabe","suffix":""},{"id":225606641,"identity":"92ac3b48-5264-403f-8484-102affe26b26","order_by":14,"name":"Gen Kondoh","email":"","orcid":"","institution":"Laboratory of Animal Experiments for Regeneration, Institute for Frontier Medical Sciences, Kyoto University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gen","middleName":"","lastName":"Kondoh","suffix":""},{"id":225606642,"identity":"240a710a-c40f-42cc-b3a8-4a160ad8de07","order_by":15,"name":"Kyoji Horie","email":"","orcid":"https://orcid.org/0000-0001-9036-8180","institution":"Nara Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Kyoji","middleName":"","lastName":"Horie","suffix":""},{"id":225606643,"identity":"7aa7c9e5-6c08-41bf-86d1-54b1e867f6ae","order_by":16,"name":"Naoki Hosen","email":"","orcid":"https://orcid.org/0000-0001-9570-0947","institution":"Osaka University Graduate School of Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Naoki","middleName":"","lastName":"Hosen","suffix":""},{"id":225606644,"identity":"3d66e755-51a7-4f3b-8ae0-32d918778988","order_by":17,"name":"Shigetoshi Sano","email":"","orcid":"https://orcid.org/0000-0002-9812-0216","institution":"Kōchi University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shigetoshi","middleName":"","lastName":"Sano","suffix":""},{"id":225606645,"identity":"fbdb647f-29c2-41d4-8697-ae8e47d1bc0d","order_by":18,"name":"Junji Takeda","email":"","orcid":"","institution":"Research Institute for Microbial Diseases, Osaka University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Junji","middleName":"","lastName":"Takeda","suffix":""}],"badges":[],"createdAt":"2022-12-31 10:55:40","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2431052/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2431052/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41467-024-49252-7","type":"published","date":"2024-06-25T04:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":41656051,"identity":"65a2333d-137a-4005-91b4-54ce4d4168e8","added_by":"auto","created_at":"2023-08-16 19:12:54","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":176383,"visible":true,"origin":"","legend":"\u003cp\u003eHomozygous mutant ESCs screening identified Ahed. a Procedure of screening homozygous mutant mouse ESC lines by deriving haematopoietic (myeloid and erythroid) cells on OP9 stromal cells. SCF, stem cell factor; EPO, erythropoietin; IL-3, interleukin 3; PECAM1, platelet/endothelial cell adhesion molecule 1. b Phase contrast images of cells on day 10 of ESC differentiation, exhibiting proliferating haematopoietic cells except for in the Ahedm/m culture. Sparkling particles observed in Ahedm/m culture are lipid droplets present in OP9-derived adipocytes (original magnification, ×200). Ahedwt/wt, parental wild-type ESCs; Ahedm/m, homozygous gene-trap mutant ESCs; Ahedm/r, heterozygous revertant ESCs; Ahedr/r, homozygous revertant ESCs. c The numbers of non-adherent (blood) cells on day 13 (n = 3); **p \u0026lt; 0.01. d Flow cytometric analysis of haematopoietic cells harvested on day 13 of culture. Percentages of cells in each quadrant are shown. Note that the elapsed time in applying each sample was set to be equal. e May-Grünwald/Giemsa-stained cytocentrifuge preparations from each culture on day 13. Scale bars, 50 μm. f The numbers of haematopoietic cells on day 13 (n = 3). EV, empty vector; cDNA, Ahed cDNA. **p \u0026lt; 0.01. g Flow cytometric analysis of haematopoietic cells harvested on day 13 of each culture. h MayGrünwald/Giemsa-stained cytocentrifuge preparations from each culture on day 13. Scale bars, 50 μm. Data in all quantitative panels are presented as mean ± SEM.\u003c/p\u003e","description":"","filename":"f1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/19a311471a79515fabaae23d.jpg"},{"id":41656833,"identity":"1b3482d2-efb5-4f77-bb25-d59e9ce06356","added_by":"auto","created_at":"2023-08-16 19:20:53","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":138434,"visible":true,"origin":"","legend":"\u003cp\u003eThe Ahed gene product is a nuclear protein. a Confocal microscopy images of Ahedm/m ESCs expressing EGFP-tagged full length Ahed cDNA. EGFP-Ahed, Ahed cDNA tagged with EGFP at the 5ʹend; Ahed-EGFP, Ahed cDNA tagged with EGFP at the 3ʹ-end. Nuclei are counterstained with Hoechst 33342. Note that EGFP fluorescence is localised in the nucleus. Scale bars, 20 μm. b Schematic of the AHED protein. Red boxes indicate K(K/R)X(K/R) sequences, which are distributed within the domain between a.a. residues 151–260. N, N-terminus; C, C-terminus. c Deletion of a.a. 151–260 resulted in EGFP fluorescence in both cytoplasm and nuclei (upper). Conversely, a.a. residues 151–260 are sufficient for nuclear localisation of the fusion protein (middle). Diffuse distribution of the EGFP protein is shown as a control (lower). Scale bars, 20 μm. d Confocal microscopy images of Ahedm/m ESCs expressing EGFP-tagged deletion mutants (upper, deletion of a.a. 151–200; lower, deletion of a.a. 201– 260). Nuclei are counterstained with Hoechst 33342. Apart from mitotic cells, EGFP fluorescence is localised in the nucleus. Red boxes in the left diagrams indicate K(K/R)X(K/R) sequences. Scale bars, 20 μm.\u003c/p\u003e","description":"","filename":"f2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/44e5eeca12ed4608e5a6d9f0.jpg"},{"id":41656049,"identity":"d730abf2-b6eb-48f1-ab6e-c9f4b582d3bb","added_by":"auto","created_at":"2023-08-16 19:12:53","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":153184,"visible":true,"origin":"","legend":"\u003cp\u003eAhed deletion causes severely anaemic and embryonic demise. a Genotyping results of the pups and embryos obtained from Ahed+/−; Vav1-cre and Ahedfl/fl pairs at the indicated points. *Chisquare test against expectations of Mendelian segregation. b Embryos recovered at E12.5. Asterisks indicate the Ahed cKO (Vav1-cre Ahedfl/fl) embryos. Scale bars, 5 mm. c Embryos recovered at E16.5 (left). Arrows indicate Ahed cKO embryos. Scale bar, 5 mm. The embryo in the dotted square exhibits subcutaneous oedema (arrowhead, right). d E14.5 foetal liver cells were stained with an isotype control IgG (dashed line) or an anti-TER119 Ab (solid line) and analysed by flow cytometry. Representative data for the indicated genotypes are shown. Absolute numbers of TER119+ erythroid cells in E14.5 foetal liver were calculated from the flow cytometric data. Horizontal lines indicate median values. **p \u0026lt; 0.01. e Graph showing the proportion of E14.5 foetal liver erythroid cells in each developmental stage. ProE, proerythroblasts; B, basophilic cells; Poly, polychromatic cells; A, acidophilic cells; R, denucleated red blood cells. *p \u0026lt; 0.05; n.s., not significant. f Flow cytometry plots of E14.5 foetal liver erythrocytes from Vav1-cre Ahedfl/+ (control) and Vav1-cre Ahedfl/fl (cKO) mice to evaluate erythroid differentiation using Ter119 and CD71. Each subset group was defined as follows; S0, Ter119- CD71- ; S1, Ter119- CD71+; S2, Ter119LoCD71+; S3, Ter119+CD71+; S4, Ter119- CD71+. Numbers indicate the percentage of each fraction. Results shown are representative of three independent experiments. Data in all quantitative panels are presented as mean ± SEM.\u003c/p\u003e","description":"","filename":"f3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/555e94add0bbbbb60dbde512.jpg"},{"id":41656052,"identity":"48ddd42e-9afd-4c1f-a91b-66737a9014f8","added_by":"auto","created_at":"2023-08-16 19:12:54","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":258606,"visible":true,"origin":"","legend":"\u003cp\u003eAhed is indispensable for HSPC differentiation. a Absolute numbers of LSK cells in E14.5 foetal liver (FL). b Absolute numbers of LT-HSCs, MPPs, LMPPs, and CLPs; n = 3–5, pooled from two independent experiments. In bar charts, results are shown as mean ± SEM and each dot represents an individual FL result. c Absolute numbers of CMPs, GMPs, MEPs; n = 2–6. d A schematic representation summarizing (a–c). e LSK cells from E14.5 FL were subjected to methylcellulose culture. Generated colonies were classified and counted. BFU-E, burst-forming units-erythroid; CFU-G/M/GM, colony-forming unit-granulocyte/macrophage/granulocyte-macrophage; *p \u0026lt; 0.05; **p \u0026lt; 0.01. f E14.5 FL LSK cells were co-cultured with MS-5 in the presence of SCF, Flt3-ligand and IL-7. Flow cytometry profiles of recovered CD45+ cells (left). Absolute numbers of CD19+ B cells differentiated from 2.5 × 102 LSK cells (right). *p \u0026lt; 0.05; **p \u0026lt; 0.01. g Limiting dilution analysis of E14.5 FL LSK cells using MS-5 coculture. Frequencies of haematopoietic progenitor cells were determined and shown in parenthesis. NA, not analysed. h Transplantation strategy. Donor Vav1-cre Ahedfl/+ (control) or Vav1-cre Ahedfl/fl (cKO) E14.5 LSK cells sorted from CD45.1- CD45.2+ and transplanted into lethally irradiated CD45.1+ CD45.2+ WT mice (n = 7–8) with 5 × 105 CD45.1+ CD45.2+ WT BM cells as a rescue. i Chimerism of peripheral blood in recipients. Numbers in each figure indicate percentages of each fraction. Results shown are representative of two independent experiments. j Chimerism of LSK in recipients’ bone marrow. Numbers in each figure indicate percentages of each fraction. Results shown are representative of two independent experiments. Data in all quantitative panels are presented as mean ± SEM.\u003c/p\u003e","description":"","filename":"f4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/2b84acb8b0456f4bed61839b.jpg"},{"id":41656835,"identity":"90db44a0-1b46-4261-a827-0f235b9d2738","added_by":"auto","created_at":"2023-08-16 19:20:54","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":145308,"visible":true,"origin":"","legend":"\u003cp\u003eAhed deficiency affects the expression of multiple genes. a Differentially expressed genes from RNA-seq in foetal liver (FL) were compared with those in adult bone marrow (BM), in which gene expressions of Vav1-cre Ahedfl/fl (cKO) versus Vav1-cre Ahedfl/+ (control) in E14.5 FL, and R26CE Ahedfl/fl (cKO) versus R26CE Ahedfl/+ (control) mice in BM were analysed. A strong positive correlation was observed between the two groups. The Venn diagram shows the number of differentially expressed genes that overlapped between the two groups. b The top 10 enriched biological functional categories in which gene expressions of cKO versus control conducted in E14.5 FL (left) and adult BM (right), according to the Ingenuity Knowledge Base. The x-axis shows the significance, which is the value of – log (p value). c Upstream transcriptional regulators in Ahed cKO FLs compared with control FL (left), and in R26CE Ahedfl/fl (cKO) BM compared with control BM (right). d Expression levels of representative haematopoietic stem cell-related genes, Gata2, Lmo2, Runx1 and Tal1 in E14.5 foetal liver LSK cells. Data are shown as mean expression levels ± standard deviation. a.u., arbitrary unit. Vav1-cre Ahedfl/+ (n = 5); Vav1-cre Ahedfl/fl (n = 6). *p \u0026lt; 0.05. Data in all quantitative panels are presented as mean ± SEM.\u003c/p\u003e","description":"","filename":"f5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/81fb4aee22fe484df80f4842.jpg"},{"id":41656834,"identity":"5012c757-e413-416d-94c6-eb677b06ee87","added_by":"auto","created_at":"2023-08-16 19:20:54","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":83456,"visible":true,"origin":"","legend":"\u003cp\u003eAhed deficiency induces cellular apoptosis. a Number of cells yielded on day 5. **p \u0026lt; 0.01. b Flow cytometry contour plots showing live (PI−, Annexin V−), early apoptotic (PI−, Annexin V+), late apoptotic (PI+, Annexin V+), and necrotic (PI+, Annexin V−) cell fractions on day 5 (left). The CD11b+ fraction was analysed. Percentages of cells in each quadrant are shown (right). **p \u0026lt; 0.01; ***p \u0026lt; 0.001. c DNA content of cultured cells on day 5 (left). The proportions of apoptotic cells (DNA content \u0026lt; 2n) and cells in S-G2/M phase are shown (right). *p \u0026lt; 0.05; ***p \u0026lt; 0.001. Data in all quantitative panels are presented as mean ± SEM.\u003c/p\u003e","description":"","filename":"f6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/0fa4289957b6a74cfddb3d67.jpg"},{"id":41656054,"identity":"a8788e11-0c34-4d19-814a-0aac9c11f4d0","added_by":"auto","created_at":"2023-08-16 19:12:54","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":244390,"visible":true,"origin":"","legend":"\u003cp\u003eAhed deficiency induces abnormal transcripts of critical genes for haematopoiesis. a Visualisation using the biophysical interactions of ORFeome-based complexes 3.0 (BioPlex 3.0) network database indicated a close correlation of the Ahed protein with numerous RNA splicing related proteins. Combined data obtained from HEK293T (human renal cancer cell line) and HCT116 (human colon cancer cells). b Splicing event visualisation in four representative genes, PU.1, Ythdf2, Sel1l, and Zfp60 involved in haematopoietic stem cells through the application of Leafcutter. c Schematic diagram of specific primer design to examine splicing and aberrant transcripts by conventional reverse transcription PCR (RT-PCR). d Electrophoresis results of RT-PCR with each specific primer (Supplementary Table 5). Results shown are representative of two genes. e Intron sequences extracted from gels of long PCR products and identified by Sanger sequencing. Result shown is representative of a gene Ythdf2. f Enriched pathways in genes (intron clusters with FDR \u0026lt; 0.05 and mapped to a single gene) by Enrichr. g List of haematopoietic neoplasm patients with C11orf57 mutations from the database (COSMIC; https://cancer.sanger.ac.uk/cosmic/gene/analysis); somatic mutations of Ahed were found in some patients with haematopoietic neoplasm such as acute myeloid leukemia, myelodysplastic syndrome, and chronic lymphocytic leukemia. h List of patients with C11orf57 mutations from the database, including splicing variant patterns (St. Jude Cloud PeCan; https://pecan.stjude.cloud/proteinpaint/)\u003c/p\u003e","description":"","filename":"f7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/14da60ccd5c198737abdeb93.jpg"},{"id":59081589,"identity":"32bcdb13-fbd3-4e7b-88c3-f6ce13c64b82","added_by":"auto","created_at":"2024-06-26 07:05:42","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1709359,"visible":true,"origin":"","legend":"","description":"","filename":"NakaiAhedArticleFile.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1_covered_96221368-2b18-4eda-b93f-79caf36f436d.pdf"},{"id":41656048,"identity":"a4ff279f-90cf-4bec-8aa3-dd26785bdc5e","added_by":"auto","created_at":"2023-08-16 19:12:53","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":15623,"visible":true,"origin":"","legend":"Dataset 1","description":"","filename":"SupplementaryData1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/bbd20d028b3f095ab35c0d91.xlsx"},{"id":41656056,"identity":"8488efc4-3d91-497b-90e6-dc8b79b4ba19","added_by":"auto","created_at":"2023-08-16 19:12:54","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":4709705,"visible":true,"origin":"","legend":"","description":"","filename":"NakaiAhedSupplementaryInformation.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2431052/v1/0aec20311bed2067491d1ff7.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"A newly identified gene Ahed plays essential roles in haematopoiesis through RNA splicing","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Ahed, haematopoiesis, mutant embryonic stem cells screening, nuclear protein, RNA splicing, C11orf57","lastPublishedDoi":"10.21203/rs.3.rs-2431052/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2431052/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The development of haematopoiesis is regulated by coordinated actions of multiple genes. Mutations in some genes are intimately involved in the occurrence of haematological cancers. Here, we report a previously undefined functional gene in haematopoiesis, which we isolated by screening mutant embryonic stem cells. The gene, named “\u003ci\u003eattenuated haematopoietic development\u003c/i\u003e” (\u003ci\u003eAhed\u003c/i\u003e), encoded an uncharacterized nuclear protein. We found that \u003ci\u003eAhed\u003c/i\u003e conditional knock out (cKO) foetuses became anaemic after E14.5 and died before birth. Transplantation experiments revealed that \u003ci\u003eAhed\u003c/i\u003e-deficient lineage\u003csup\u003e-\u003c/sup\u003eSca-1\u003csup\u003e+\u003c/sup\u003ec-Kit\u003csup\u003eHi\u003c/sup\u003e cells were unable to reconstitute haematopoiesis \u003ci\u003ein vivo\u003c/i\u003e, suggesting that \u003ci\u003eAhed\u003c/i\u003e is indispensable for functional haematopoietic stem/progenitor cells. Multiple RNA sequencing analyses of \u003ci\u003eAhed\u003c/i\u003e-cKO cells revealed that this protein governs various critical pathways by regulating alternative splicing of RNAs. Published databases of cancer-related mutations present blood cancer patients with somatic mutations in the \u003ci\u003eAhed\u003c/i\u003e gene. Thus, \u003ci\u003eAhed\u003c/i\u003e plays a fundamental role in haematopoiesis through RNA splicing, whose mutations relates to malignancies.","manuscriptTitle":"A newly identified gene Ahed plays essential roles in haematopoiesis through RNA splicing","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-08-16 19:12:49","doi":"10.21203/rs.3.rs-2431052/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"8762111a-a0f1-4623-bed8-e65f71774b47","owner":[],"postedDate":"August 16th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":23964535,"name":"Biological sciences/Developmental biology/Haematopoiesis/Lymphopoiesis"},{"id":23964536,"name":"Biological sciences/Genetics/Development"},{"id":23964537,"name":"Biological sciences/Stem cells/Haematopoietic stem cells"},{"id":23964538,"name":"Biological sciences/Genetics/RNA splicing"},{"id":23964539,"name":"Biological sciences/Stem cells/Embryonic stem cells"}],"tags":[],"updatedAt":"2024-06-26T07:05:35+00:00","versionOfRecord":{"articleIdentity":"rs-2431052","link":"https://doi.org/10.1038/s41467-024-49252-7","journal":{"identity":"nature-communications","isVorOnly":false,"title":"Nature Communications"},"publishedOn":"2024-06-25 04:00:00","publishedOnDateReadable":"June 25th, 2024"},"versionCreatedAt":"2023-08-16 19:12:49","video":"","vorDoi":"10.1038/s41467-024-49252-7","vorDoiUrl":"https://doi.org/10.1038/s41467-024-49252-7","workflowStages":[]},"version":"v1","identity":"rs-2431052","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2431052","identity":"rs-2431052","version":["v1"]},"buildId":"GqpaHPwrfC8PjnIFayRh5","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}