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All Journal Farmasains : Jurnal Farmasi dan Ilmu Kesehatan Jurnal Ilmiah Farmasi Statistika Jurnal Kedokteran Brawijaya Jurnal Teknosains Indonesian Journal of Cancer Chemoprevention Indonesian Journal of Pharmaceutical Science and Technology STOMATOGNATIC- Jurnal Kedokteran Gigi Indonesian Journal of Biotechnology Journal of Agromedicine and Medical Sciences Pharmacon Majalah Obat Tradisional Journal of Tropical Biodiversity and Biotechnology INDONESIAN JOURNAL OF PHARMACY JURNAL BIOLOGI INDONESIA Dental Journal (Majalah Kedokteran Gigi) The Indonesian Biomedical Journal Padjadjaran Journal of Dentistry Jurnal Kimia Terapan Indonesia JFIOnline YARSI Medical Journal Majalah Kesehatan Pharmamedika Pharmaceutical Sciences and Research (PSR)
Edy Meiyanto
Faculty Of Pharmacy, Universitas Gadjah Mada
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Civil Engineering, Building, Construction & Architecture Computer Science & IT Decision Sciences, Operations Research & Management Dentistry Economics, Econometrics & Finance Environmental Science Health Professions Immunology & microbiology Industrial & Manufacturing Engineering Materials Science & Nanotechnology Mathematics Mechanical Engineering Medicine & Pharmacology Public Health

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Journal : Jurnal Teknosains

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Optimized condition for pei-based transient transfection of lifeact-gfp/nls-mcherry expressing plasmid used as cell barcode for syncytia live cell imaging Kumara, Dennaya; Harsan, Hayfa Salsabila; Novianti, Metta; Lestari, Dinda; Septisetyani, Endah Puji; Prasetyaningrum, Pekik Wiji; Paramitasari, Komang Alit; Meiyanto, Edy
Jurnal Teknosains Vol 13, No 1 (2023): December
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/teknosains.89479

Abstract

The transfection efficiency positively affects the successful plasmid DNA transfer into cells, with the highlight on the amount of plasmid DNA and its ratio to the transfection reagent. Polyethyleneimine (PEI) is a cost-effective transfection reagent that facilitates DNA transfer by forming positively charged DNA complexes. It allows DNA to interact with negatively charged cell surfaces and enter the cells by endocytosis. In this study, we optimized the condition for transient transfection of life act-GFP/NLS-mCherry-expressing plasmid in BHK-21 and 293T cells using PEI. This plasmid is helpful as a biosensor of the cytoskeleton and nucleus that enables live imaging observation using a fluorescence microscope, for instance, in the observation of syncytium. Here, we optimized two independent variables: the amount of DNA (0.5 and 1 µg) and the ratio of DNA-PEI (1:3 and 1:4). GFP and mCherry expressions were observed at 24, 48, and 72 h post-transfection. As a result, transfection efficiency achieved by using PEI in 293T cells is higher than in BHK-21 cells, which are ~90% and ~50%, respectively. Moreover, amongst four different transfection conditions, in both cell lines, 1 µg of plasmid DNA with a 1:3 DNA-PEI ratio yields the most efficiency with the least amount of toxicity. We used this condition for the syncytia observation in 293T cells as a model of the cell-to-cell transmission of SARS-CoV-2. Syncytia formation was successfully observed by detecting the giant cells expressing GFP/mCherry with multiple nuclei.
Co-Authors . Anindyajati . Larasati . Sugiyanto Adam Hermawan Adam Hermawan Adisusilo, Midori Rahmadhany Putri Aditya Fitriasari Aditya Fitriasari Agung Endro Nugroho Agusta Fauzi, Ilham Agustina Setiawati Ainun Wulandari Alexxander, . Alexxander, . Ameilinda Monikawati Ameilinda Monikawati Andita Pra Darma Andita Pra Darma Andriyani, Rina Angelina, Marissa Angraini, Sonia Meta Anif Nur Artanti, Anif Nur Ardriyanto, Maria Indra Arief Nurrochmad Arief Rahman Hakim Asih Triastuti Astrid Ayu Maruti Aulia Katarina Aulia Rahman, Faaza Ayu Maruti, Astrid B. Sudarto Banun Kusumawardani Chio Oka, Chio D., Andita Pra D., Andita Pra Da'i, Muhammad Da’i, Muhammad Dewi Arum Sekti, Dewi Arum Dewi Pamungkas Putri, Dyaningtyas Dewi Pratiwi Dini Maharani Djaswadi Dasuki Dwi Ana Nawangsari Dwi Ana Nawangsari, Dwi Ana Dwi Merry Christmarini Robin Dwi Nurahmanto Dyaningtyas D. P. Putri Dyaningtyas Dewi Putri Pamungkas Effendi, Fatiha Citra Endah Puji Septisetyani, Endah Puji Endah Puspitasari Endang Lukitaningsih Endang Purwaningsih Erlina Rivanti Erna Prawita Setyowati Esti, Yuni Fajar Fany Mutia Cahyani Farmasyanti, Cendrawasih Andusyana Feby Handoko, Fransiscus Fikri Amalia Fina Aryani Goenadi Fina Aryani Goenadi, Fina Aryani Fitria Rahmi Fiveri, Anis Fiveri, Anis Fortunella Tjondro Handayani, Sri Handayani, Sri Hanifa, Mila Hapsari, Novia Permata Hargiani, Fransisca Xaveria Harsan, Hayfa Salsabila Hastuti, Hanaan Emilia Adi Hendra Kurniawan Maury Hendri Wasito Heni Susilowati Herwandhani Putri Herwandhani Putri Ibrahim Arifin Ida Ayu Putu Sri Widnyani Ika Nurzijah Ika Rahmawati Sutejo Ikawati, Muthi' Ilham Agusta Fauzi Ilham Agusta Fauzi Imono Argo Donatus, Imono Argo Indrasetiawan, Puguh Indri Kusharyanti Inna Armandani, Inna Inna Armandari Iwan Sahrial Hamid JAKA WIDADA Jauhari, Fahmi Ihsanuddin Jenie, Riris Istighfari Jenie, Riris Istighfari Juni Ekowati Kadarsih Soejono, Sri Kadarsih Soejono, Sri Kartika Dyah Palupi Kartika Dyah Palupi Kato, Jun-Ya Kawai, Taro Kholid Alfan Nur Kholid Alfan Nur Komang Alit Paramitasari Kuijpers-Jagtman, Anne Marie Kumara, Dennaya Laras Widawaty Putri Lestari, Dinda Luthfia Indriyani M, Kawaichi M, Kawaichi Mae Sri Hartati Wahyuningsih Maran, Gergorius Gena Marcellino Rudyanto Maria Dwi Supriyati Maria Dwi Supriyati, Maria Dwi Masashi Kawaichi Masashi Kawaichi, Masashi Muflikhasari, Haruma Anggraini Muhammad Da'i Muhammad Fithrul Mubarok, Muhammad Fithrul Muthi Ikawati N., Perdana Adhi N., Perdana Adhi Nabila, Klarissa Nanda Resa Pratama Niken Nur W, Niken Novi Hastuti, Novi Novianti, Metta Novitasari, Dhania Nugraheni, Nadzifa Nunuk Aries Nurulita Nunuk Purwanti Nurma Sabila Nurrachma, Marsya Yonna P.K.W., Diah Ayu P.K.W., Diah Ayu Perdana Adhi Nugroho Perdana Adhi Nugroho Prasetyaningrum, Pekik Wiji Pudjono Pudjono Putri, Amaliya Permata Putri, Herwandhani Putri, Nindya Budiana R A Susidarti Raditya Prima Istiaji Rahmah, Inggita Hasi Rahman, Faaza Aulia Rahmawati, Desty Restia Rahmi Khamsita Ramadani, Ratna Dwi Ratih Hardika Pratama Ratna Asmah Susidarti Ratna Asmah Susidarti Retno Ardhani Retno Murwanti Rifai, Fauziah Novita Putri Riris I Jenie Riris I Jenie, Riris I Riris Istighfari Jenie Riris Istighfari Jenie Riris Istighfari Jenie Risdian, Chandra Rita Riata Rohmad Yudi Utomo Rohmad Yudi Utomo Rosa Adelina Rosana Anna Ashari Rosana Anna Ashari, Rosana Anna Rosye H.R. Tanjung Rul Afiyah Syarif Rumiyati, Rumiyati Salsabila, Irfani Aura Santoso, Christopher Filando Sari Haryanti Sari Haryanti Sarmoko Sarmoko Sendy Junedi Sendy Junedy, Sendy Shigeru Sasaki Sismindari . Sitarina Widyarini Sofa Farida Sri . Handayani Sri Handayani Sri Kadarsih Soejono Sri Kasianningsih Sri Susilowati Sri Tasminatun Sugeng Riyanto Sugiyanto . Sugiyanto . Sukardiman Supardjan A. M. Supardjan A.M., Supardjan Supardjan AM Supardjan AM, Supardjan Susi Ari Kristina Tutuk Budiati Udin, Zalinar Umar A. Jenie Umar Anggara Jenie Umar Anggara Jenie Umar Anggara Jenie Widayanti, Wasita Rachma Yundari, Yundari Yurista Gilang Yurista Gilang Ikhtiarsyah Yuyun Farida Yuyun Farida, Yuyun Zufairo, Shofa Khamdanatuz Zulfin, Ummi Maryam