A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium Discoideum
The social amoeba Dictyostelium discoideum is a commonly used eukaryotic model organism for the study of cell division, chemotaxis, differentiation, phagocytosis, and other cellular processes. Electroporation is an effective and efficient method for delivering plasmid DNA into D. discoideum, an inva...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Western Libraries, The University of Western Ontario
2023-05-01
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| Series: | Journal of Open Hardware |
| Subjects: | |
| Online Access: | https://account.openhardware.metajnl.com/index.php/up-j-joh/article/view/52 |
| _version_ | 1827199500898271232 |
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| author | Michael C. Cauchy Ali A. Khan Yulia Artemenko David A. Dunn |
| author_facet | Michael C. Cauchy Ali A. Khan Yulia Artemenko David A. Dunn |
| author_sort | Michael C. Cauchy |
| collection | DOAJ |
| description | The social amoeba Dictyostelium discoideum is a commonly used eukaryotic model organism for the study of cell division, chemotaxis, differentiation, phagocytosis, and other cellular processes. Electroporation is an effective and efficient method for delivering plasmid DNA into D. discoideum, an invaluable tool for studying intracellular processes. The technology is readily available but often prohibitively expensive. Although several custom-built electroporation devices have been developed, none deliver the specific 8.5kV/cm exponentially decaying waveform required for D. discoideum transformation. The present study examined whether a simple, inexpensive device can be built to produce this waveform through a simple resistor-capacitor (RC) circuit. A pulse generator RC circuit was built incorporating inexpensive electronic components and a 3D printed cuvette chamber. All four possible combinations of custom-built and commercial pulse generators and custom-built and commercial cuvette chambers were used to transform D. discoideum cells with a plasmid encoding green fluorescent protein (GFP). There were no significant differences in the number of surviving cells immediately following or 24 hours post-transformation between the systems. All combinations of custom-built and commercial systems achieved comparably high transformation efficiency shown by percent of cells expressing GFP six days after the transformation. Since the waveform-specific electroporation system we present here can be built by non-experts with easily obtainable materials and 3D printing, we envision this device to benefit investigators in areas with low research budgets and educators in multiple STEM fields. |
| first_indexed | 2024-03-13T05:36:12Z |
| format | Article |
| id | doaj.art-0bd0c065c78643dc8c74d2209e1c7b50 |
| institution | Directory Open Access Journal |
| issn | 2514-1708 |
| language | English |
| last_indexed | 2025-03-21T10:33:54Z |
| publishDate | 2023-05-01 |
| publisher | Western Libraries, The University of Western Ontario |
| record_format | Article |
| series | Journal of Open Hardware |
| spelling | doaj.art-0bd0c065c78643dc8c74d2209e1c7b502024-07-02T22:04:47ZengWestern Libraries, The University of Western OntarioJournal of Open Hardware2514-17082023-05-01715510.5334/joh.523A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium DiscoideumMichael C. Cauchy0https://orcid.org/0000-0001-7085-7788Ali A. Khan1https://orcid.org/0000-0003-0337-6735Yulia Artemenko2https://orcid.org/0000-0001-7456-8924David A. Dunn3https://orcid.org/0000-0001-9275-4798Dept. of Biological Sciences at the State University of New York at Oswego, Oswego, NYDept. of Biological Sciences at the State University of New York at Oswego, Oswego, NYDept. of Biological Sciences at the State University of New York at Oswego, Oswego, NYDept. of Biological Sciences at the State University of New York at Oswego, Oswego, NYThe social amoeba Dictyostelium discoideum is a commonly used eukaryotic model organism for the study of cell division, chemotaxis, differentiation, phagocytosis, and other cellular processes. Electroporation is an effective and efficient method for delivering plasmid DNA into D. discoideum, an invaluable tool for studying intracellular processes. The technology is readily available but often prohibitively expensive. Although several custom-built electroporation devices have been developed, none deliver the specific 8.5kV/cm exponentially decaying waveform required for D. discoideum transformation. The present study examined whether a simple, inexpensive device can be built to produce this waveform through a simple resistor-capacitor (RC) circuit. A pulse generator RC circuit was built incorporating inexpensive electronic components and a 3D printed cuvette chamber. All four possible combinations of custom-built and commercial pulse generators and custom-built and commercial cuvette chambers were used to transform D. discoideum cells with a plasmid encoding green fluorescent protein (GFP). There were no significant differences in the number of surviving cells immediately following or 24 hours post-transformation between the systems. All combinations of custom-built and commercial systems achieved comparably high transformation efficiency shown by percent of cells expressing GFP six days after the transformation. Since the waveform-specific electroporation system we present here can be built by non-experts with easily obtainable materials and 3D printing, we envision this device to benefit investigators in areas with low research budgets and educators in multiple STEM fields.https://account.openhardware.metajnl.com/index.php/up-j-joh/article/view/523d printinggenetic engineeringgmoopen-sourcetransformation |
| spellingShingle | Michael C. Cauchy Ali A. Khan Yulia Artemenko David A. Dunn A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium Discoideum Journal of Open Hardware 3d printing genetic engineering gmo open-source transformation |
| title | A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium Discoideum |
| title_full | A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium Discoideum |
| title_fullStr | A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium Discoideum |
| title_full_unstemmed | A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium Discoideum |
| title_short | A Low-Cost Electroporator for Genetically Modifying Social Amoeba Dictyostelium Discoideum |
| title_sort | low cost electroporator for genetically modifying social amoeba dictyostelium discoideum |
| topic | 3d printing genetic engineering gmo open-source transformation |
| url | https://account.openhardware.metajnl.com/index.php/up-j-joh/article/view/52 |
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