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利用 CRISPR/Cas 基因组编辑技术进行大麦育种的研究进展。

CRISPR/Cas genome editing perspectives for barley breeding.

机构信息

Laboratory of Cell Engineering, Institute of Plant Biology and Biotechnology, Almaty, Kazakhstan.

出版信息

Physiol Plant. 2022 May;174(3):e13686. doi: 10.1111/ppl.13686.

Abstract

The CRISPR/Cas9 technology shows potential to improve crop breeding efficiency and antiviral defense. The interest in DNA editing in crops has grown due to the possibility of increasing the resistance of different plants to many viruses. Our aim was to create an elite disease-resistant local barley cultivar using CRISPR/Cas9 biotechnology. For this purpose, we used CRISPR/Cas 9-eIF4E with the eukaryotic translation initiation factor 4E (eIF4E) barley gene to edit the genomes of five local Kazakhstan barley cultivars. After identifying the single guide RNA (sgRNA) target sequences, they were synthesized and cloned into the CRISPR-plant vector before being introduced into barley cells via our own patented Agrobacterium germ-line transformation technique. Barley plants eIF4E-modified were successfully obtained and were resistant to virus infection. Based on our research, the CRISPR/Cas9 system for plant genome editing could be a prospect for applying this breakthrough biotechnology in barley breeding.

摘要

CRISPR/Cas9 技术有望提高作物育种效率和抗病毒防御能力。由于有可能提高不同植物对许多病毒的抗性,因此人们对作物中的 DNA 编辑产生了兴趣。我们的目的是利用 CRISPR/Cas9 生物技术创建一个具有优良抗病性的当地大麦品种。为此,我们使用了 CRISPR/Cas9-eIF4E 和真核翻译起始因子 4E(eIF4E)大麦基因来编辑五个哈萨克斯坦当地大麦品种的基因组。在确定了单指导 RNA(sgRNA)的靶序列后,我们将它们合成并克隆到 CRISPR-plant 载体中,然后通过我们自己的专利农杆菌生殖系转化技术将其导入大麦细胞。成功获得了 eIF4E 修饰的大麦植物,并对病毒感染具有抗性。基于我们的研究,植物基因组编辑的 CRISPR/Cas9 系统可能是将这项突破性生物技术应用于大麦育种的前景。

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