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

FEBS Letters

article

Rupam Kumar Bhunia¹  Guillaume N. Menard¹  Peter J. Eastmond¹ 

[1] Plant Sciences and the Bioeconomy, Rothamsted Research;National Agri-Food Biotechnology Institute

关键词: CRISPR/Cas9;    genome editing;    promoter fusion;    transcriptional gain-of-function;    transgene-free;    plant oil metabolism;   

DOI  :  10.1002/1873-3468.14365

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown