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.
引用
收藏
页数:8
相关论文
共 63 条
[21]   Digital Microfluidic Approach for Efficient Electroporation with High Productivity: Transgene Expression of Microalgae without Cell Wall Removal [J].
Im, Do Jin ;
Jeong, Su-Nam ;
Yoo, Byeong Sun ;
Kim, Bolam ;
Kim, Dong-Pyo ;
Jeong, Won-Joong ;
Kang, In Seok .
ANALYTICAL CHEMISTRY, 2015, 87 (13) :6592-6599
[23]   Discrete Electrostatic Charge Transfer by the Electrophoresis of a Charged Droplet in a Dielectric Liquid [J].
Im, Do Jin ;
Ahn, Myung Mo ;
Yoo, Byeong Sun ;
Moon, Dustin ;
Lee, Dong Woog ;
Kang, In Seok .
LANGMUIR, 2012, 28 (32) :11656-11661
[24]   Electrophoresis of a Charged Droplet in a Dielectric Liquid for Droplet Actuation [J].
Im, Do Jin ;
Noh, Jihoon ;
Moon, Dustin ;
Kang, In Seok .
ANALYTICAL CHEMISTRY, 2011, 83 (13) :5168-5174
[25]   Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding [J].
Jarrell, Justin A. ;
Twite, Amy A. ;
Lau, Katherine H. W. J. ;
Kashani, Moein N. ;
Lievano, Adrian A. ;
Acevedo, Julyana ;
Priest, Craig ;
Nieva, Jorge ;
Gottlieb, David ;
Pawell, Ryan S. .
SCIENTIFIC REPORTS, 2019, 9
[26]   Advances in the delivery of RNA therapeutics: from concept to clinical reality [J].
Kaczmarek, James C. ;
Kowalski, Piotr S. ;
Anderson, Daniel G. .
GENOME MEDICINE, 2017, 9
[27]   Exosomes facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer [J].
Kamerkar, Sushrut ;
LeBleu, Valerie S. ;
Sugimoto, Hikaru ;
Yang, Sujuan ;
Ruivo, Carolina F. ;
Melo, Sonia A. ;
Lee, J. Jack ;
Kalluri, Raghu .
NATURE, 2017, 546 (7659) :498-+
[28]   Genome-wide measurement of RNA secondary structure in yeast [J].
Kertesz, Michael ;
Wan, Yue ;
Mazor, Elad ;
Rinn, John L. ;
Nutter, Robert C. ;
Chang, Howard Y. ;
Segal, Eran .
NATURE, 2010, 467 (7311) :103-107
[29]   Insulin-like growth factor 1 (IGF1), IGF binding protein 3 (IGFBP3), and breast cancer risk: pooled individual data analysis of 17 prospective studies [J].
Key, Timothy J. ;
Appleby, Paul N. ;
Reeves, Gillian K. ;
Roddam, Andrew W. .
LANCET ONCOLOGY, 2010, 11 (06) :530-542
[30]   Effects of Actin Cytoskeleton Disruption on Electroporation In Vitro [J].
Kim, Hong Bae ;
Lee, Seho ;
Chung, Jong Hoon ;
Kim, Seong Nam ;
Sung, Chang Kyu ;
Baik, Ku Youn .
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 2020, 191 (04) :1545-1561