舌下腺黏液性腺泡细胞特异性Cre重组酶基因敲入小鼠的建立和鉴定OACSTPCD
Generation of a Cre recombinase knock-in mouse line labeling the mucous acinar cells of the sublingual gland
目的 建立和鉴定整合素β样蛋白1基因(Itgbl1)启动子指导下的Cre重组酶基因敲入小鼠.方法 利用CRISPR/Cas9基因编辑技术研制Itgbl1-Cre基因敲入小鼠.利用Itgbl1-Cre基因敲入小鼠与Cre的报道小鼠ROSALSL-tdTomato交配的子代小鼠进行遗传细胞谱系示踪.通过红色荧光蛋白tdTomato鉴定Itgbl1阳性细胞及其子代细胞的组织表达谱.通过多色免疫荧光染色利用多种细胞特异性的标志物鉴定阳性细胞及其子代细胞的细胞类型.结果与结论 tdTomato蛋白在舌下腺的黏液性腺泡细胞、胰岛细胞和胃的分泌细胞中特异性表达.此外,tdTomato在部分视网膜和脑的神经元细胞以及少量肠的浆膜层、骨关节软骨、骨膜和骨髓的细胞中表达.成功建立了首个在小鼠舌下腺的黏液性腺泡细胞中特异性表达Cre重组酶的转基因小鼠品系.
Objective To generate and identify the Itgbl1(integrin beta-like)promoter-driven Cre knock-in mouse line.Methods Itgbll-Cre knock-in mice were generated using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing.The Itgbl1-Cre mice were crossed with the Cre reporter ROSALSL-tdTomato)mice to detect the expression profile of Cre activity.The tdTomato expression pattern across tissues and cell-specific markers were used to identify the cell types of Itgbl1-expressing cells and their progeny.Results and Conclusion tdTomato was specifically expressed in mucous acinar cells of the sublingual gland,pancreatic islet cells,and gastric endocrine cells.In addition,tdTomato expression was also found in some of the neurons of the retina and brain,as well as in a few cells in the serosal layer of the intestine,articular cartilage,periosteum,and bone marrow.The first Itgbl1-Cre recombinase transgenic mouse line was established,which can specifically label the mucous acinar cells of the sublingual gland.
彭艳丽;杨冠;滕艳;粟柯糙;郎依铭;谢中良;李明月;周学涛;王庆业;汪海珍;杨晓
军事科学院军事医学研究院,国家蛋白质科学中心(北京),北京 102206
生物学
整合素β样蛋白1Cre重组酶遗传细胞谱系示踪多色免疫荧光
integrin beta-like 1Cre recombinasegenetic cell lineage tracingmulticolor immunofluorescence
《军事医学》 2024 (006)
429-433 / 5
国家重点研发计划(2018YFA0801104);国家自然科学基金面上项目(82372721)
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