Condition optimization for electroporation transfection in horse skeletal muscle satellite cells.

IF 1.7 3区农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Animal Biotechnology Pub Date : 2024-11-01 Epub Date: 2023-11-20 DOI:10.1080/10495398.2023.2280664

Tseweendolmaa Ulaangerel, Minna Yi, Undarmaa Budsuren, Yingchao Shen, Hong Ren, Bold Demuul, Dongyi Bai, Dulguun Dorjgotov, Gantulga Davaakhuu, Tuyatsetseg Jambal, Manglai Dugarjav, Gerelchimeg Bou

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Abstract

Satellite cells are an important cellular model for studying muscle growth and development and mammalian locomotion-related molecular mechanisms. In this study, we investigated the effects of voltage, pulse duration, and DNA dosage on horse skeletal muscle satellite cells' electroporation transfection efficiency using the eukaryotic expression plasmid Td Tomato-C1 (5.5 kb) encoding the red fluorescent protein gene mainly based on fluorescence-positive cell rate and cell survival rate. By comparison of different voltages, pulse durations, and DNA doses, horse skeletal muscle satellite cells have nearly 80% transfection efficiency under the condition of voltage 120 V, DNA dosage 7 µg/ml, and pulse duration 30 ms. This optimized electroporation condition would facilitate the application of horse skeletal muscle satellite cells in genetic studies of muscle function and related diseases.

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马骨骼肌卫星细胞电穿孔转染条件优化。

卫星细胞是研究肌肉生长发育和哺乳动物运动相关分子机制的重要细胞模型。本研究以编码红色荧光蛋白基因的真核表达质粒Td Tomato-C1 (5.5 kb)为载体，主要基于荧光阳性细胞率和细胞存活率，研究了电压、脉冲持续时间和DNA剂量对马骨骼肌卫星细胞电穿孔转染效率的影响。通过对不同电压、脉冲时间和DNA剂量的比较，在电压120 V、DNA剂量7µg/ml、脉冲时间30 ms条件下，马骨骼肌卫星细胞转染效率接近80%。优化后的电穿孔条件将促进马骨骼肌卫星细胞在肌肉功能和相关疾病的遗传学研究中的应用。

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来源期刊

Animal Biotechnology 工程技术-奶制品与动物科学

CiteScore

2.90

自引率

5.40%

发文量

230

审稿时长

>12 weeks

期刊介绍： Biotechnology can be defined as any technique that uses living organisms (or parts of organisms like cells, genes, proteins) to make or modify products, to improve plants, animals or microorganisms for a specific use. Animal Biotechnology publishes research on the identification and manipulation of genes and their products, stressing applications in domesticated animals. The journal publishes full-length articles and short research communications, as well as comprehensive reviews. The journal also provides a forum for regulatory or scientific issues related to cell and molecular biology applied to animal biotechnology. Submissions on the following topics are particularly welcome: - Applied microbiology, immunogenetics and antibiotic resistance - Genome engineering and animal models - Comparative genomics - Gene editing and CRISPRs - Reproductive biotechnologies - Synthetic biology and design of new genomes