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Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2

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...

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Autores principales: Zhao, Lihua, Li, Meishuang, Yin, Zhibao, Lv, Limin, Zhou, Meng, Wang, Yixi, Zhang, Manling, Guo, Tianxu, Guo, Xiyun, Liu, Han, Cheng, Linxin, Liang, Xiubin, Duo, Shuguang, Li, Rongfeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9740088/
https://www.ncbi.nlm.nih.gov/pubmed/36497134
http://dx.doi.org/10.3390/cells11233874
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author Zhao, Lihua
Li, Meishuang
Yin, Zhibao
Lv, Limin
Zhou, Meng
Wang, Yixi
Zhang, Manling
Guo, Tianxu
Guo, Xiyun
Liu, Han
Cheng, Linxin
Liang, Xiubin
Duo, Shuguang
Li, Rongfeng
author_facet Zhao, Lihua
Li, Meishuang
Yin, Zhibao
Lv, Limin
Zhou, Meng
Wang, Yixi
Zhang, Manling
Guo, Tianxu
Guo, Xiyun
Liu, Han
Cheng, Linxin
Liang, Xiubin
Duo, Shuguang
Li, Rongfeng
author_sort Zhao, Lihua
collection PubMed
description 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.
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spelling pubmed-97400882022-12-11 Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2 Zhao, Lihua Li, Meishuang Yin, Zhibao Lv, Limin Zhou, Meng Wang, Yixi Zhang, Manling Guo, Tianxu Guo, Xiyun Liu, Han Cheng, Linxin Liang, Xiubin Duo, Shuguang Li, Rongfeng Cells Article 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. MDPI 2022-12-01 /pmc/articles/PMC9740088/ /pubmed/36497134 http://dx.doi.org/10.3390/cells11233874 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhao, Lihua
Li, Meishuang
Yin, Zhibao
Lv, Limin
Zhou, Meng
Wang, Yixi
Zhang, Manling
Guo, Tianxu
Guo, Xiyun
Liu, Han
Cheng, Linxin
Liang, Xiubin
Duo, Shuguang
Li, Rongfeng
Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2
title Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2
title_full Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2
title_fullStr Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2
title_full_unstemmed Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2
title_short Development of a Lung Vacancy Mouse Model through CRISPR/Cas9-Mediated Deletion of Thyroid Transcription Factor 1 Exon 2
title_sort development of a lung vacancy mouse model through crispr/cas9-mediated deletion of thyroid transcription factor 1 exon 2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9740088/
https://www.ncbi.nlm.nih.gov/pubmed/36497134
http://dx.doi.org/10.3390/cells11233874
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