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High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction
Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631631/ https://www.ncbi.nlm.nih.gov/pubmed/31185602 http://dx.doi.org/10.3390/mi10060387 |
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author | Zhang, Jianxiong Hu, Yawei Wang, Xiaoqing Liu, Peng Chen, Xiaofang |
author_facet | Zhang, Jianxiong Hu, Yawei Wang, Xiaoqing Liu, Peng Chen, Xiaofang |
author_sort | Zhang, Jianxiong |
collection | PubMed |
description | Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, and transduction can be conducted efficiently on our previously developed superhydrophobic microwell array chip (SMAR-chip). A total of 169 chemical transfections were successfully performed on the chip in physically separated microwells through a few simple steps, contributing to the convenience of DNA delivery and media change on the SMAR-chip. Efficiencies comparable to the traditional transfection in multi-well plates (~65%) were achieved while the manual operations were largely reduced. Two transfection procedures, the dry method amenable for the long term storage of the transfection material and the wet method for higher efficiencies were developed. Multiple transfections in a scheduled manner were performed to further increase the transfection efficiencies or deliver multiple genes at different time points. In addition, high throughput virus packaging integrated with target cell transduction were also proved which resulted in a transgene expression efficiency of >70% in NIH 3T3 cells. In summary, the SMAR-chip based high throughput gene delivery is efficient and versatile, which can be used for large scale genetic modifications in a variety of cell types. |
format | Online Article Text |
id | pubmed-6631631 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-66316312019-08-19 High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction Zhang, Jianxiong Hu, Yawei Wang, Xiaoqing Liu, Peng Chen, Xiaofang Micromachines (Basel) Article Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, and transduction can be conducted efficiently on our previously developed superhydrophobic microwell array chip (SMAR-chip). A total of 169 chemical transfections were successfully performed on the chip in physically separated microwells through a few simple steps, contributing to the convenience of DNA delivery and media change on the SMAR-chip. Efficiencies comparable to the traditional transfection in multi-well plates (~65%) were achieved while the manual operations were largely reduced. Two transfection procedures, the dry method amenable for the long term storage of the transfection material and the wet method for higher efficiencies were developed. Multiple transfections in a scheduled manner were performed to further increase the transfection efficiencies or deliver multiple genes at different time points. In addition, high throughput virus packaging integrated with target cell transduction were also proved which resulted in a transgene expression efficiency of >70% in NIH 3T3 cells. In summary, the SMAR-chip based high throughput gene delivery is efficient and versatile, which can be used for large scale genetic modifications in a variety of cell types. MDPI 2019-06-10 /pmc/articles/PMC6631631/ /pubmed/31185602 http://dx.doi.org/10.3390/mi10060387 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Zhang, Jianxiong Hu, Yawei Wang, Xiaoqing Liu, Peng Chen, Xiaofang High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction |
title | High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction |
title_full | High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction |
title_fullStr | High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction |
title_full_unstemmed | High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction |
title_short | High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction |
title_sort | high-throughput platform for efficient chemical transfection, virus packaging, and transduction |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631631/ https://www.ncbi.nlm.nih.gov/pubmed/31185602 http://dx.doi.org/10.3390/mi10060387 |
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