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Generation of transgenic fish cell line with a-lactalbumin nanocarriers co-delivering Tol2 transposase mRNA and plasmids

文献类型： 外文期刊

作者： Zhao, Ran 1 ; Zhang, Yan 1 ; Wang, Qi 1 ; Cao, Yi-Ming 1 ; Hou, Ming-Xi 1 ; Sun, Xiao-Qing 1 ; Yu, Shuang-Ting 1 ; Chen, Ying-Jie 4 ; Wang, Kai-Kuo 4 ; Li, Jiong-Tang 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Key Lab Aquat Genom, Minist Agr & Rural Affairs, Beijing 100141, Peoples R China

2.Chinese Acad Fishery Sci, Beijing Key Lab Fishery Biotechnol, Beijing 100141, Peoples R China

3.Chinese Acad Agr Sci, Beijing 100081, Peoples R China

4.Shanghai Ocean Univ, Natl Demonstrat Ctr Expt Fisheries Sci Educ, Shanghai 201306, Peoples R China

期刊名称：ISCIENCE （ 影响因子：4.6； 五年影响因子：5.0 ）

ISSN：

年卷期： 2024 年 27 卷 8 期

页码：

收录情况： SCI

摘要： Fish cells, such as grass carp ( Ctenopharyngodon idella) ) kidney (CIK) cells, are harder to transfect than mammalian cells. There is a need for an efficient gene delivery system for fish cells. Here, we used CIK cell line as a model to develop a strategy to enhance RNA and plasmid DNA transfection efficiency using a nanocarrier generated from cc-lactalbumin ( cc-NC). cc-NC absorbed nucleic acid cargo efficiently and exhibited low cytotoxicity. Plasmid transfection was more efficient with cc-NC than with liposomal transfection reagents. We used cc-NC to co-transfect Tol2 transposase mRNA and a plasmid containing Cas9 and GFP, generating a stable transgenic CIK cell line. Genome and RNA sequencing revealed that the Cas9 and GFP fragments were successfully inserted into the genome of CIK cells and efficiently transcribed. In this study, we established an efficient transfection system for fish cells using cc-NC, simplifying the process of generating stable transgenic fish cell lines.

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