CRISPR/Cas9介导的基因组缺失产生的天然启动子-基因融合提供了一种无转基因的方法来驱动叶片中的油脂积累

IF 3 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

FEBS Letters Pub Date : 2022-05-01 DOI:10.1002/1873-3468.14365

R. Bhunia, Guillaume N. Menard, P. Eastmond

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引用次数: 8

摘要

在不插入外源DNA的情况下实现功能获得表型是植物生物技术学家面临的一项重要挑战。在这里，我们表明，通过使用双引导CRISPR/Cas9删除干预基因组序列，可以将基因置于上游基因启动子的控制之下。我们将一个非必需光合作用相关基因的启动子与拟南芥脂肪酶缺乏糖依赖性1突变体中的二甘醇酰基转移酶2（DGAT2）融合以驱动叶片中的异位油积累。DGAT2的表达增强了20倍以上，三酰甘油含量增加了约30倍。这种缺失策略提供了一条无转基因的途径来改造依赖转录功能获得的性状，例如生产高脂饲料以提高反刍动物养殖的生产力和可持续性。

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A native promoter–gene fusion created by CRISPR/Cas9‐mediated genomic deletion offers a transgene‐free method to drive oil accumulation in leaves

Achieving gain‐of‐function phenotypes without inserting foreign DNA is an important challenge for plant biotechnologists. Here, we show that a gene can be brought under the control of a promoter from an upstream gene by deleting the intervening genomic sequence using dual‐guide CRISPR/Cas9. We fuse the promoter of a nonessential photosynthesis‐related gene to DIACYLGLYCEROL ACYLTRANSFERASE 2 (DGAT2) in the lipase‐deficient sugar‐dependent 1 mutant of Arabidopsis thaliana to drive ectopic oil accumulation in leaves. DGAT2 expression is enhanced more than 20‐fold and the triacylglycerol content increases by around 30‐fold. This deletion strategy offers a transgene‐free route to engineering traits that rely on transcriptional gain‐of‐function, such as producing high lipid forage to increase the productivity and sustainability of ruminant farming.

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来源期刊

FEBS Letters 生物-生化与分子生物学

CiteScore

6.60

自引率

2.90%

发文量

303

审稿时长

1 months

期刊介绍： FEBS Letters is one of the world''s leading journals in molecular biology and is renowned both for its quality of content and speed of production. Bringing together the most important developments in the molecular biosciences, FEBS Letters provides an international forum for Minireviews, Research Letters and Hypotheses that merit urgent publication.