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电转笔:一种超低成本、无电源、便携式电穿孔仪。

ElectroPen: An ultra-low-cost, electricity-free, portable electroporator.

机构信息

School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, United States of America.

Lambert High School, Suwanee, Georgia, United States of America.

出版信息

PLoS Biol. 2020 Jan 10;18(1):e3000589. doi: 10.1371/journal.pbio.3000589. eCollection 2020 Jan.

DOI:10.1371/journal.pbio.3000589
PMID:31922526
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6953602/
Abstract

Electroporation is a basic yet powerful method for delivering small molecules (RNA, DNA, drugs) across cell membranes by application of an electrical field. It is used for many diverse applications, from genetically engineering cells to drug- and DNA-based vaccine delivery. Despite this broad utility, the high cost of electroporators can keep this approach out of reach for many budget-conscious laboratories. To address this need, we develop a simple, inexpensive, and handheld electroporator inspired by and derived from a common household piezoelectric stove lighter. The proposed "ElectroPen" device can cost as little as 23 cents (US dollars) to manufacture, is portable (weighs 13 g and requires no electricity), can be easily fabricated using 3D printing, and delivers repeatable exponentially decaying pulses of about 2,000 V in 5 ms. We provide a proof-of-concept demonstration by genetically transforming plasmids into Escherichia coli cells, showing transformation efficiency comparable to commercial devices, but at a fraction of the cost. We also demonstrate the potential for rapid dissemination of this approach, with multiple research groups across the globe validating the ease of construction and functionality of our device, supporting the potential for democratization of science through frugal tools. Thus, the simplicity, accessibility, and affordability of our device holds potential for making modern synthetic biology accessible in high school, community, and resource-poor laboratories.

摘要

电穿孔是一种将小分子(RNA、DNA、药物)穿过细胞膜的基本而强大的方法,通过施加电场来实现。它被用于许多不同的应用,从基因工程细胞到药物和 DNA 疫苗的输送。尽管有如此广泛的用途,但电穿孔仪的高成本可能使许多注重预算的实验室无法使用这种方法。为了解决这个问题,我们受一种常见的家用压电打火机的启发,开发了一种简单、廉价、手持的电穿孔仪。该设备的制造成本可低至 23 美分(美元),便携(重 13 克,无需电力),可以使用 3D 打印轻松制造,并且可以提供重复的、指数衰减的约 2000V 的脉冲,持续时间为 5ms。我们通过将质粒基因转化为大肠杆菌细胞提供了一个概念验证演示,结果表明转化效率与商用设备相当,但成本却大大降低。我们还展示了这种方法快速传播的潜力,全球多个研究小组验证了我们设备的构建和功能的易用性,这支持了通过节俭工具实现科学民主化的潜力。因此,我们的设备简单、易用、价格低廉,有可能使现代合成生物学在高中、社区和资源匮乏的实验室中得到普及。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/540ce652e552/pbio.3000589.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/5d9c3f86b894/pbio.3000589.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/e0ba4716e49f/pbio.3000589.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/05cbefb3c1c7/pbio.3000589.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/540ce652e552/pbio.3000589.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/5d9c3f86b894/pbio.3000589.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/e0ba4716e49f/pbio.3000589.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/05cbefb3c1c7/pbio.3000589.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e1/6953602/540ce652e552/pbio.3000589.g004.jpg

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