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| 1 | TRDMT1 exhibited protective effects against LPS-induced inflammation in rats through TLR4-NF-κB/MAPK-TNF-αpathway显示文摘Background:Inflammation is a complex physiological and pathological process.Although many types of inflammation are well characterized,their physiological func-tions are largely unknown.tRNA aspartic acid methyltransferase 1(TRDMT1)has been implicated as a stress-related protein,but its intrinsic biological role is unclear.Methods:We constructed a Trdmt1 knockout rat and adopted the LPS-induced sepsis model.Survival curve,histopathological examination,expression of inflammatory fac-tors,and protein level of TLR4 pathway were analyzed.Results:Trdmt1 deletion had no obvious impact on development and growth.Trdmt1 de-letion slightly increased the mortality during aging.Our data showed that Trdmt1 strongly responded in LPS-treated rats,and Trdmt1 knockout rats were vulnerable to LPS treat-ment with declined survival rate.We also observed more aggravated tissue damage and more cumulative functional cell degeneration in LPS-treated knockout rats compared with control rats.Further studies showed upregulated TNF-αlevel in liver,spleen,lung,and serum tissues,which may be explained by enhanced p65 and p38 phosphorylation.Conclusions:Our data demonstrated that Trdmt1 plays a protective role in inflamma-tion by regulating the TLR4-NF-κB/MAPK-TNF-αpathway.This work provides useful information to understand the TRDMT1 function in inflammation. | Zhengguang Li Xiaolong Qi Xu Zhang Lei Yu Lijuan Gao Weining Kong Wei Chen Wei Dong Lijun Luo Dan Lu Lianfeng Zhang Yuanwu Ma | 2022 | Animal Models and Experimental Medicine2022，5，2： | 5 |
| 2 | 肌肉特异性表达Cas9示踪同源打靶载体的构建及其在C2C12细胞中的整合显示文摘目的本研究拟构建肌肉特异表达Cas9示踪同源打靶载体,为成肌细胞分化研究及肌肉特异表达Cas9小鼠模型制作奠定基础。方法人工合成肌肉特异启动子SP,替换PX459载体中的CMV启动子,构建肌肉特异表达Cas9载体;载体在C2C12细胞中的编辑效率由XbaI和T7E1酶切检测;在此基础上通过同源重组引入DsRed红色荧光蛋白构建Cas9示踪载体;将示踪载体的SP-Cas9-DsRed部分连接至Rosa26位点左右同源臂之间,构建肌肉特异表达Cas9示踪重组载体;将该载体与PX459-Rosa26共转染至C2C12细胞并进行嘌呤霉素筛选,观察荧光的表达,同时,提取细胞DNA进行PCR鉴定和测序,检测载体在C2C12细胞中的整合情况。结果酶切鉴定以及对目的片段的测序结果表明,成功构建了肌肉特异表达Cas9载体PX459-Rosa26-SP和肌肉特异同源打靶示踪载体Donor-Cas9-SP-DsRed;XbaI和T7E1酶切实验表明,PX459-Rosa26-SP在C2C12细胞中具有较高的编辑效率;转染后的C2C12细胞出现红色荧光,表明Donor-Cas9-SP-DsRed具有表达活性且在肌肉细胞中特异表达;PCR鉴定和测序结果表明,Donor-Cas9-SP-DsRed在C2C12细胞Rosa26位点成功整合。结论成功构建了肌肉特异表达Cas9示踪同源打靶载体Donor-Cas9-SP-DsRed,在为肌肉特异表达Cas9小鼠模型制备提供载体基础的同时,也为肌肉相关基因疾病的研究与基因治疗提供了新的思路。 | 王晓萌 周慧敏 董奕彤 陈胜男 安铁洙 殷萍 王春生 | 2022 | 中国实验动物学报2022，30，3： | 1 |
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