دورية أكاديمية
Nucleofection as an Efficient Method for Alpha TC1-6 Cell Line Transfection
| العنوان: | Nucleofection as an Efficient Method for Alpha TC1-6 Cell Line Transfection |
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| المؤلفون: | Marija Đorđević, Verica Paunović, Maja Jovanović Tucović, Anja Tolić, Jovana Rajić, Svetlana Dinić, Aleksandra Uskoković, Nevena Grdović, Mirjana Mihailović, Ivanka Marković, Jelena Arambašić Jovanović, Melita Vidaković |
| المصدر: | Applied Sciences, Vol 12, Iss 15, p 7938 (2022) |
| بيانات النشر: | MDPI AG, 2022. |
| سنة النشر: | 2022 |
| المجموعة: | LCC:Technology LCC:Engineering (General). Civil engineering (General) LCC:Biology (General) LCC:Physics LCC:Chemistry |
| مصطلحات موضوعية: | alpha TC1-6 cells, transfection, PEI, nucleofection, green fluorescent protein, EpiCRISPR, Technology, Engineering (General). Civil engineering (General), TA1-2040, Biology (General), QH301-705.5, Physics, QC1-999, Chemistry, QD1-999 |
| الوصف: | An efficient transfection is a crucial step for the introduction of epigenetic modification in host cells, and there is a need for an optimized transfection process for individual model systems separately. Mouse pancreatic αTC1-6 cells, which act as an attractive model system for epigenetic cell reprogramming and diabetes treatment, were transiently transfected with two different transfection methods: the chemical method with polyethyleneimine (PEI) and nucleofection as a physical transfection method. Flow cytometry and fluorescent microscopy examination of GFP expression showed that transfection efficiency was affected by the size of plasmids using both transfection methods. Subsequently, the Cas9 mRNA expression confirmed successful transfection with EpiCRISPR plasmid, whereas the cell physiology remained unchanged. The adjusted nucleofection protocol for αTC1-6 cells transfected with an EpiCRISPR mix of plasmids reached 71.1% of GFP-positive transfected cells on the fifth post-transfection day and proved to be much more efficient than the 3.8% GFP-positive PEI transfected cells. Modifying the protocol, we finally specify CM-156 program and SF 4D-Nucleofector X Solutions for Amaxa™ nucleofection as a method of choice for alpha TC1-6 cell line transfection. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2076-3417 |
| العلاقة: | https://www.mdpi.com/2076-3417/12/15/7938Test; https://doaj.org/toc/2076-3417Test |
| DOI: | 10.3390/app12157938 |
| الوصول الحر: | https://doaj.org/article/16c02b491d4948f28ad24661373c371bTest |
| رقم الانضمام: | edsdoj.16c02b491d4948f28ad24661373c371b |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 20763417 |
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| DOI: | 10.3390/app12157938 |