Metode CRISPR/CAS dan Minimalisasi Off-Target: Review
DOI:
https://doi.org/10.61761/agiotech.1.1.17-30Keywords:
CRISPR, Cas9, sgRNA, Off-targetAbstract
Aplikasi bioteknologi dengan pendekatan rekayasa genetika seperti genome editing berhasil dalam memperbaiki sifat tanaman dengan tujuan meningkatkan kualitas hasil. Keberhasilan CRISPR/Cas9 untuk memodifikasi genom dengan presisi yang belum pernah terjadi sebelumnya dapat disebabkan oleh akurasi, efisiensi, efektivitas biaya, dan kemudahan penggunaan. Mekanisme pengeditan genom CRISPR/Cas9 adalah memanipulasi gen dengan cara menyisipkan, mengganti, menghapus satu basa atau lebih pada sekuen tertentu. Oleh karena adanya penyisipan atau pergantian gen tersebut, metode CRISPR/Cas dapat menyebabkan efek di luar target (off-target) yang merusak pada tingkat genomik. Artikel ini membahas perkembangan CRISPR/Cas9, mekanisme CRISPR/Cas9, dan metode minimalisasi off-target yang sering terjadi pada sistem CRISPR/Cas9. Metode dalam meminimalisir off-target dari penggunaan CRISPR/Cas9 dapat dilakukan melalui beberapa cara yaitu: (1) modifikasi sgRNA terkait konten GC sgRNA, panjang sgRNA, truncated gRNA, modifikasi kimia sgRNA, dan modifikasi sgRNA in silico, (2) modifikasi protein Cas, dan (3) metode delivery CRISPR
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Copyright (c) 2023 Sophia Sophia, Muhammad Antony, Muh Aswad Ashan, Hafizh Fadhullah, Rika M Jannah
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