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All Journal Tropical Medicine Journal Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences International Journal of Biosciences and Biotechnology Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Jurnal Kesehatan Masyarakat Indonesian Journal of Urology Jurnal Farmasi Klinik Indonesia Indonesian Journal of Biotechnology Jurnal Kedokteran BERITA BIOLOGI Majalah Obstetri dan Ginekologi The Indonesian Biomedical Journal Jurnal Teknologi Laboratorium Universa Medicina Paediatrica Indonesiana JURNAL MANAJEMEN DAN PELAYANAN FARMASI (Journal of Management and Pharmacy Practice) Jurnal Gizi Klinik Indonesia Majalah Farmaseutik Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal Media Ilmu Kesehatan Jurnal Kesehatan Prima Acta Biochimica Indonesiana Indonesian Journal of Clinical Pathology and Medical Laboratory (IJCPML) Journal of Community Empowerment for Health Pena Medika : Jurnal Kesehatan SWARNA Medika Respati : Jurnal Ilmiah Kesehatan Jurnal Manusia dan Lingkungan SWARNA: Jurnal Pengabdian Kepada Masyarakat
Ahmad Hamim Sadewa
Departemen Biokimia, Fakultas Kedokteran, Kesehatan Masyarakat, Dan Keperawatan Universitas Gadjah Mada, Yogyakarta, Indonesia
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Journal : The Indonesian Biomedical Journal

 1 
GSTM1 Deletion Compensated in mRNA Expression and 4T1 Viability After Editing Using CRISPR/Cas9 Single and Double gRNA Emilia Vivi Arsita; Dwi Aris Agung Nugrahaningsih; Ahmad Hamim Sadewa
The Indonesian Biomedical Journal Vol 15, No 2 (2023)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v15i2.2131

Abstract

BACKGROUND: Glutathione S-transferase Mu-1 (GSTM1) is known to undergo polymorphism and plays role in drug metabolism including Paclitaxel (PTX), the first-line chemotherapy for breast cancer. However, the effect of GSTM1 polymorphism against chemotherapy in breast cancer is limited and unexplored. This study was conducted to explore the effects of single and double guide (gRNA) on the GSTM1 knocked out (KO) and its effect on the response of PTX in the 4T1 cell line.METHODS: The preparatory stage was done by culturing and electroporating 4T1 cells using Ribonucleoprotein of clustered regularly interspaced short palindromic repeats (CRISPR)/Caspase 9 (Cas9). KO validation was examined by quantitative reverse transcription polymerase chain reaction (qRT-PCR), Sanger sequencing, and ICE analysis. The 4T1 viability was examined by MTT Assay.RESULTS: The number of base pairs of GSTM1 after being engineered by single or double gRNA was 86 bases. The DNA quantity of GSTM1 engineered by gRNA was more than using double gRNAs. The mRNA expression of GSTM1 engineered by single gRNA was lower than using double gRNAs. IC50 values of PTX between wildtype and KO were not significantly different, in the range of 30 µM.CONCLUSION: The base-pair length of GSTM1 exon 4 that is knocked out with single and double gRNA have the same number of base pairs. The quantity of GSTM1 DNA and mRNA expression are contrary between single gRNA and double gRNA, and IC50 PTX values in the 4T1 cell line of the control group with single or double gRNA knocked out do not differ markedly. PTX efficiency as chemotherapy is not disturbed in the GSTM1 deletion genetic profile.KEYWORDS: GSTM1, gRNA, Paclitaxel, CRISPR, breast cancer
Co-Authors . Harapan . HERNAYANTI . Syahrul Abdul Salam M Sofro, Abdul Salam Abdul Salam M. Sofro Abdul Salam Mudzakir Sofro Abdul Salam Mudzakir Sofro, Abdul Salam Mudzakir Abdurahman Laqif Abdurahman Laqif Abdus Samik Wahab Abrory Agus Cahya Pramana Ahmad Husain Asdie Ahmad Husain Asdie Ahmad Husain Asdie Ajeng Viska Icanervilia Akhmad Kharis Nugroho Alfasunu, Serafim Anggelia Puspasari Anggoro Budi Hartopo Ani Kristiyani Arif Faisal Arif Faisal Arta Farmawati Arta Farmawati Atmaja, Sarah Awal Prasetyo Bai Apris Bambang Hariono Bambang Hariono Bambang Hariono Bansai Immanuel Bernadia Branitamahisi Bernadia Branitamahisi Budi Yuli Setianto Cahyono Hadi Cahyono Hadi Christine Patramurti Citra Maharani Danarsih, Dwi Eni Darojatun Ida Demas Bayu Handika Dessy Rakhmawati Emril Dewajani Purnomosari Dian Caturini Sulistyonigrum Dian Lestari, Nova Dianandha Septiana Rubi Dibyo Pramono Didik Setyo Heriyanto Djaswadi Dasuki Djaswadi Dasuki Djaswadi Dasuki DONO INDARTO Dono Indarto Dono Indarto Dwi Aris Agung Nugrahaningsih Dyah Wulan Anggrahini Ellsya Angeline Rawar Emilia Vivi Arsita Emy Huriyati Endang Mutiawati Rahayuningsih* Faisal Rahman Fransisca Shinta Maharini Hanafi, Arif Riswahyudi Hari Kusnanto Hasan Sjahrir Hemi Sinorita Hendi Wicaksono Hernayanti , Hernayanti HERNAYANTI HERNAYANTI Hernayanti Hernayanti Hexa Apriliana Hidayah I Gusti Ayu Nyoman Danuyanti I NYOMAN MANTIK ASTAWA Ida Ayu Preharsini Ida Ayu Preharsini Kusuma Ika Rahayu Ika Setyawati Ikhsan, M. Robikhul Imelda, Priscillia Indwiani Astuti Indwiani Astuti Indwiani Astuti Inna Narayani Iskandar Zakaria Jayusman, Achmad Mulawarman Jenny Hidayat Jontari Hutagalung Kik Hao Samuel Kris Herawan Timotius Kusumadewi Eka Damayanti Kusumadewi Eka Damayanti Kusumadewi Eka Damayanti Lina Choridah Linawati Hananta, Linawati Lucia Krisdinarti Lukman Hakim M. Robikhul Ikhsan Maliyah Madiyan Maria Dara Novi Handayani, Maria Dara Novi Marsetyawan HNE Soesatyo Mega Tyas Prihatin Mohammad Robikhul Ikhsan Mus, Rosdiana Mustofa Mustofa Mustofa Mustofa Ngadikun - Ngadikun Ngadikun Nor Istiqomah Nor Istiqomah Nor Istiqomah Nor Sri Inayati Nor Sri Inayati Novijanti Rintis Nurtjahjo Dwi Sasongko P. Purwono Pinda Hutajulu Pinda Hutajulu, Pinda Pramudji Hastuti Pramudji Hastuti Pramudji Hastuti Pramudji Hastuti Pramudji Hastuti Pramudji Hastuti Pramudji Hastuti Pramudji Hastuti, Pramudji Prasetyastuti Prasetyastuti Prasetyastuti Prasetyastuti Prasetyastuti Prasetyastuti, . Rasmaya Niruri Ratih Feraritra Danu Atmaja Rina Pratiwi Pudja I. A Rina Susilowati RIZKI FAJAR UTAMI Rochadi Rochadi Rochadi Rochadi, Rochadi S. Supargiyono Sabirin, Rahmaningsih Mara Sarah Safira Umarghanies Saryono Saryono Satyagraha Ari Winasti Seto Priyambodo Setyo Purwono Siti Boedina Kresno Siti Wahyuningsih Sofia Mubarika Haryana Sri Sutarni Sri Wahyuni Subagus Wahyuono Subagus Wahyuono Sukarti Moeljopawiro Sukarti Moeljopawiro Sukarti Moeljopawiro Sunarti Sunarti Sunarti Sunarti Sunarti Sunarti Sunarti Sunarti, Sunarti Sunarto Ang Tasmini - Tasmini Tasmini Teguh Aryandono Titiek Suhardi Haripurnomo Kushadiwijaya Hidayati Totok Utoro Totok Utoro Triwibowo A. Garjito Umarghanies, Sarah Safira Viren Ramadhan Vitarani Dwi Ananda Ningrum Vitria Sari Dewi Vitria Sari Dewi Waode Astria Sahrani Wasilah Rochmah wayan T Artama wayan T Artama Wayan Tunas Artama Widhiastuti, Stefani Santi Wiryatun Lestariana Wiryatun Lestariana, Wiryatun Wiwik Handayani Yuliana Heri Soesilo Yuliana Heri Soesilo Yuliana Heri Soesilo Yuliani, Fara Silvia Yunilistiaingsih Yunilistiaingsih Yunilistiaingsih, Yunilistiaingsih Yuningtyaswari Yuningtyaswari Zainal Arifin Nang Agus Zullies Ikawati