BİTKİ ISLAHINDA CRISPR/CAS9 UYGULAMALARI

Year 2020, Volume: 3 Issue: 2, 32 - 40, 02.11.2020

Anıl Mehmet Baltacı Mehmet Arslan

Abstract

Bitkiler için genom dizilerinin mevcudiyeti ve genom düzenleme teknolojisindeki ilerlemeler, hemen hemen her türlü istenilen özellik açısından ıslah olanaklarını artırmıştır. ZFN (Zinc Finger Nucleas) ve TALEN (Transcription Activator-Like Effector Nuclease) gibi genom düzenleme teknolojilerindeki gelişmeler moleküler düzeyde ilgilenilen herhangi bir genin düzenlenmesini mümkün kılmıştır. Bunların aksine CRISPR / Cas9 genom düzenleme yöntemi basit tasarım ve kolay klonlama yöntemlerini içermektedir. Cas9 genomdaki birden fazla bölgeyi hedefleyen farklı kılavuz (guide) RNA'lar ile farklı birden fazla gen bölgesine müdahale edilebilmektedir. CRISPR-Cas9 modülünde hedef özgüllüğünü geliştirmek ve hedef dışı bölünmeyi azaltmak birkaç farklı modifiye Cas9 kaseti kullanılmaktadır. Ayrıca farklı bakteri türlerinden elde edilen Cas9 enzimlerinin mevcudiyeti gen düzenleme yöntemlerinin özgüllüğünü ve verimliliğini arttırmak için yeni seçenekler sunmaktadır. Bu çalışmada, bitki ıslahçılarının CRISPR / Cas9 temelli genom düzenleme araçlarını kullanarak moleküler düzyde bitki ıslahı için mevcut durumu özetlemekte ve CRISPR / Cas9'un biyotik ve abiyotik stres toleransını artırmak için kullanıldığı çalışmaları sunmaktadır.

Keywords

TALEN, ZFN, biyotik stres, abiyotik stres, bitki ıslahı, CRISPR

CRISPR / Cas9 Applications In Plant Breeding

Abstract

The availability of genome sequences for plants and advances in genome editing technology have increased breeding possibilities for almost all kinds of desired traits. Advances in genome editing technologies such as ZFN (Zinc Finger Nucleas) and TALEN (Transcription Activator-Like Effector Nuclease) have made it possible to edit any gene of interest at the molecular level. On the contrary, the CRISPR / Cas9 genome editing method includes simple design and easy cloning methods. With different guide RNAs targeting more than one region in the Cas9 genome, multiple different gene regions can be intervened. Several different modified Cas9 cassettes are used in the CRISPR-Cas9 module to improve target specificity and reduce off-target cleavage. In addition, the availability of Cas9 enzymes from different bacterial species offers new options to increase the specificity and efficiency of gene editing methods. This study summarizes the current situation for plant breeding at the molecular level using the genome editing tools of plant breeders based on CRISPR / Cas9 and presents studies using CRISPR / Cas9 to increase biotic and abiotic stress tolerance.

Keywords

CRISPR, TALEN, ZFN, biotic stress, abiotic stress, plant breeding

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