Safe and efficient RNA and DNA introduction into cells using digital electroporation system

被引:0
作者
Bae, Seo Jun [1 ]
Im, Do Jin [1 ]
机构
[1] Pukyong Natl Univ, Dept Chem Engn, 45 Yongso Ro, Pusan 48513, South Korea
基金
新加坡国家研究基金会;
关键词
RNA delivery; Electroporation; K562; Chlamydomoans reinhardtii; Delivery system; SECONDARY STRUCTURE; CHARGED DROPLET; GROWTH; ELECTROPHORESIS; INSULIN; QUANTIFICATION; OPTIMIZATION; TRANSFECTION; EXPRESSION; DELIVERY;
D O I
10.1016/j.bioelechem.2022.108268
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We systematically compared the delivery and expression efficiencies according to cell types (plant and animal cells) and genetic materials (RNA and DNA) to deliver RNA using a digital electroporation system. Despite the significantly lower RNA delivery in Chlamydomoans reinhartii than DNA delivery due to RNA secondary structure and cell wall, the expression/delivery ratio of RNA was significantly higher than that of DNA (up to 90%), confirming the generally known fact that RNA is more favorable for expression than DNA. On the other hand, in K562 cells, the difference in RNA and DNA delivery efficiency was negligible. Therefore, structural differences between DNA and RNA affect delivery efficiency differently depending on the cell type. RNA delivery efficiency of K562 cells was high, but expression efficiency was much lower than that of microalgae. According to the proposed strategy, compatibility between K562 cells and the nucleic acids used in this study is presumed to be one of the reasons for this low expression efficiency. Gene regulation by delivering small interfering RNA (siRNA) was demonstrated in K562 cells, confirming the feasibility of the digital electroporation system for RNA inter-ference (RNAi) research as a safe and efficient delivery system.
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页数:8
相关论文
共 63 条
[51]   CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future [J].
Uddin, Fathema ;
Rudin, Charles M. ;
Sen, Triparna .
FRONTIERS IN ONCOLOGY, 2020, 10
[52]   The delivery challenge: fulfilling the promise of therapeutic genome editing [J].
van Haasteren, Joost ;
Li, Jie ;
Scheideler, Olivia J. ;
Murthy, Niren ;
Schaffer, David, V .
NATURE BIOTECHNOLOGY, 2020, 38 (07) :845-855
[53]   A Pliable Electroporation Patch (ep-Patch) for Efficient Delivery of Nucleic Acid Molecules into Animal Tissues with Irregular Surface Shapes [J].
Wei, Zewen ;
Huang, Yuanyu ;
Zhao, Deyao ;
Hu, Zhiyuan ;
Li, Zhihong ;
Liang, Zicai .
SCIENTIFIC REPORTS, 2015, 5
[54]   TUFM-knockdown inhibits the migration and proliferation of gastrointestinal stromal tumor cells [J].
Weng, Xiaoyuan ;
Zheng, Song ;
Shui, Hanli ;
Lin, Guosheng ;
Zhou, Yongjian .
ONCOLOGY LETTERS, 2020, 20 (05)
[55]   The challenge and prospect of mRNA therapeutics landscape [J].
Weng, Yuhua ;
Li, Chunhui ;
Yang, Tongren ;
Hu, Bo ;
Zhang, Mengjie ;
Guo, Shuai ;
Xiao, Haihua ;
Liang, Xing-Jie ;
Huang, Yuanyu .
BIOTECHNOLOGY ADVANCES, 2020, 40
[56]   RNA structure from deep sequencing [J].
Westhof, Eric .
NATURE BIOTECHNOLOGY, 2015, 33 (09) :928-929
[57]   Delivery Aspects of CRISPR/Cas for in Vivo Genome Editing [J].
Wilbie, Danny ;
Walther, Johanna ;
Mastrobattista, Enrico .
ACCOUNTS OF CHEMICAL RESEARCH, 2019, 52 (06) :1555-1564
[58]  
Wybranietz WA, 1999, J GENE MED, V1, P265
[59]   An improved method for circular RNA purification using RNase R that efficiently removes linear RNAs containing G-quadruplexes or structured 3′ ends [J].
Xiao, Mei-Sheng ;
Wilusz, Jeremy E. .
NUCLEIC ACIDS RESEARCH, 2019, 47 (16) :8755-8769
[60]   Extracellular vesicles as a platform for membrane-associated therapeutic protein delivery [J].
Yang, Yoosoo ;
Hong, Yeonsun ;
Cho, Eunji ;
Kim, Gi Beom ;
Kim, In-San .
JOURNAL OF EXTRACELLULAR VESICLES, 2018, 7 (01)