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All Journal AGRIEKONOMIKA Jurnal Agribisnis Indonesia (Journal of Indonesian Agribusiness) Forum Agribisnis Mimbar Agribisnis: Jurnal Pemikiran Masyarakat Ilmiah Berwawasan Agribisnis AGROMIX Jurnal Jamu Indonesia Jurnal Ilmiah Edunomika (JIE) Jurnal ABDINUS : Jurnal Pengabdian Nusantara Jurnal Bisnis Tani Journal of Integrated Agribusiness Abdi Laksana : Jurnal Pengabdian Kepada Masyarakat Community Development Journal: Jurnal Pengabdian Masyarakat IJGIE (International Journal of Graduate of Islamic Education) AGRISCIENCE Jurnal Sains Pertanian Agridev Journal : Agribusiness Digitalization & Community Development Agrimics Journal Halal Studies and Society Indonesian Journal of Jamu Journal of Empowerment Indonesia Biodiversitas Journal of Biological Diversity
Ifan Rizky Kurniyanto
Program Studi Agribisnis, Fakultas Pertanian, Universitas Trunojoyo Madura
Religion Agriculture, Biological Sciences & Forestry Humanities Biochemistry, Genetics & Molecular Biology Chemistry Computer Science & IT Decision Sciences, Operations Research & Management Economics, Econometrics & Finance Education Environmental Science Health Professions Industrial & Manufacturing Engineering Law, Crime, Criminology & Criminal Justice Medicine & Pharmacology Nursing Public Health Social Sciences Veterinary Other

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Construction of a CRISPR/Cas9-mediated genome editing system in manipulating OsART1 from Oryza sativa cv. Inpago 5 MIFTAHUL HUDA FENDIYANTO; EKO SETIAWAN; MENTARI PUTRI PRATAMI; IFAN RIZKY KURNIYANTO; FANDRI SOFIANA FASTANTI
Biodiversitas Journal of Biological Diversity Vol. 26 No. 2 (2025)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d260241

Abstract

Abstract. Fendiyanto MH, Setiawan E, Pratami MP, Kurniyanto IR, Fastanti FS. 2025. Construction of a CRISPR/Cas9-mediated genome editing system in manipulating OsART1 from Oryza sativa cv. Inpago 5. Biodiversitas 26: 920-927. Understanding the mechanism of aluminum (Al) tolerance in rice (Oryza sativa) cv. Inpago 5 can be done by mutating the ART1 gene to see which Al tolerance genes have decreased expression globally. This study aimed to construct a pRGEB32 vector containing sgRNA-ART1 derived from rice cv. Inpago 5. The construction of the pRGEB32-sgART1 vector was carried out by designing sgRNA, restriction-ligation, transformation, plasmid isolation, verification of target inserts, and sequencing of sgRNA-ART1. The results indicate that the construction of the recombinant vector pRGEB32-sgRNAART1 was carried out by combining digestion and ligase reactions. The construction of the recombinant pRGEB32 construction showed a high level of success in this study. The success of the ART1 sgRNA amplicon in recombinant Escherichia coli indicated this. Annotation studies showed that the pRGEB32 cassette plasmid sequence showed more than 99% homology with the recombinant pRGEB32-sgART1 sequence, especially only 1% was different, namely 20 bp of sgRNA that appeared different. After checking with alignment on the plasmid annotation using the MAAFT alignment method, it was found that 20 bp of ART1 sgRNA was successfully inserted into the pRGEB32-sgART1 recombinant vector. In conclusion, the construction of pRGEB32-sgART1 in this study was successful and showed the presence of a 425 bp band amplicon for the insertion from the region between M13 reverse and sgRNA-ART1.
Co-Authors Afriani, Ferina Andari, Laila Ayu Andini Dhamayanti Arfiyanti, Nurul Arifiyanti, Nurul Arifiyanti, Nurul Arinuha Novia Putri RD Azza Nuha Nisrina Badrut Tamam Bayu Krisnamurthi Destiarni, Resti Prastika Dewi, Devina Liza Shinta Dian Eswin Wijayanti Dwi Martha Yulia Dwi Ratna Hidayati EKO SETIAWAN Endang Supriatna FANDRI SOFIANA FASTANTI Fuad Hasan Fuad Hasan Husna, Harir Mamluatul Isdiana Suprapti LIA AMALIA LILIS SUAIBAH Lilis Suaibah Lukman Ismail M. Fathur Rohim Arrozy Maulida, Mitha Rachma MENTARI PUTRI PRATAMI MIFTAHUL HUDA FENDIYANTO Mochamad Aziz, Roikhan Moh. Wahyudi Priyanto Mokh Rum Mokh. Rum Mokh. Rum Muhammad Ronaydi Muhammad Syahid Musyafak Nita Alfiyani Nor Qomariyah Nurul Arifiyanti Nurul Arifiyanti Nurwijayanti Pelawi, Natasya Br Pratama, Okka Putra Pratiwi, Yunita Eka Dyah Priyanto, Moh. Wahyudi Pujiati, Tri Qoiroh, Shirotul Nur Rabbani, Bintang Cahya Rahmadilla M Gozenda Rahman Dwi Irvanto Rahmania, Handizah Kingkin Rakhamawati Riyanti Isaskar rizky, Ifan Rohim Arrozy, M. Fathur Rohman, Abdur Roikhan Mochamad Aziz, Roikhan Mochamad Romadloni, Masqotul Imam Ronaydi, Muhammad Saroh, Nisa May Setiani Setiani Setiani Setiani, Setiani Silviana Putri Slamet Subari Sriyono, Sriyono Syafruddin Syafruddin Tsaniya Rusyda Nugrahani Ulfa Utami Yudhi Prasetya Mada Yudhi Prasetya Mada Zulkhaeriyah, Zulkhaeriyah