{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T04:47:41Z","timestamp":1760244461526,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2022,12,1]],"date-time":"2022-12-01T00:00:00Z","timestamp":1669852800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Scientific Foundation of China","doi-asserted-by":"publisher","award":["31971379","KFJ-BRP-005"],"award-info":[{"award-number":["31971379","KFJ-BRP-005"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Biological Resources Program of Chinese Academy of Sciences","award":["31971379","KFJ-BRP-005"],"award-info":[{"award-number":["31971379","KFJ-BRP-005"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>A developmental niche vacancy in host embryos is necessary for stem cell complementation-based organ regeneration (SCOG). Thyroid transcription factor 1 (TTF-1) is a tissue-specific transcription factor that regulates the embryonic development and differentiation of the thyroid and, more importantly, lungs; thus, it has been considered as a master gene to knockout in order to develop a lung vacancy host. TTF-1 knockout mice were originally produced by inserting a stop codon in Exon 3 of the gene (E3stop) through embryonic stem cell-based homologous recombination. The main problems of utilizing E3stop host embryos for lung SCOG are that these animals all have a tracheoesophageal fistula (TEF), which cannot be corrected by donor stem cells, and most of them have monolateral sac-like lungs. To improve the mouse model towards achieving SCOG-based lung generation, in this project, we used the CRISPR\/Cas9 tool to remove Exon 2 of the gene by zygote microinjection and successfully produced TTF-1 knockout (E2del) mice. Similar to E3stop, E2del mice are birth-lethal due to retarded lung development with sac-like lungs and only a rudimentary bronchial tree, increased basal cells but without alveolar type II cells and blood vessels, and abnormal thyroid development. Unlike E3stop, 57% of the E2del embryos presented type I tracheal agenesis (TA, a kind of human congenital malformation) with a shortened trachea and clear separations of the trachea and esophagus, while the remaining 43% had TEF. Furthermore, all the E2del mice had bilateral sac-like lungs. Both TA and bilateral sac-like lungs are preferred in SCOG. Our work presents a new strategy for producing SCOG host embryos that may be useful for lung regeneration.<\/jats:p>","DOI":"10.3390\/cells11233874","type":"journal-article","created":{"date-parts":[[2022,12,1]],"date-time":"2022-12-01T05:03:18Z","timestamp":1669870998000},"page":"3874","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Lung Vacancy Mouse Model through CRISPR\/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2"],"prefix":"10.3390","volume":"11","author":[{"given":"Lihua","family":"Zhao","sequence":"first","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"},{"name":"Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meishuang","family":"Li","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhibao","family":"Yin","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Limin","family":"Lv","sequence":"additional","affiliation":[{"name":"Laboratory Animal Center, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meng","family":"Zhou","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9705-7964","authenticated-orcid":false,"given":"Yixi","family":"Wang","sequence":"additional","affiliation":[{"name":"Laboratory Animal Center, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manling","family":"Zhang","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tianxu","family":"Guo","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiyun","family":"Guo","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Han","family":"Liu","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Linxin","family":"Cheng","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9169-0390","authenticated-orcid":false,"given":"Xiubin","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Pathophysiology, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuguang","family":"Duo","sequence":"additional","affiliation":[{"name":"Laboratory Animal Center, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9624-3706","authenticated-orcid":false,"given":"Rongfeng","family":"Li","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing 211166, China"},{"name":"Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4528","DOI":"10.1073\/pnas.90.10.4528","article-title":"RAG-2-deficient blastocyst complementation: An assay of gene function in lymphocyte development","volume":"90","author":"Chen","year":"1993","journal-title":"Proc. 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