A native promoter–gene fusion created by CRISPR/Cas9‐mediated genomic deletion offers a transgene‐free method to drive oil accumulation in leaves. Issue 15 (6th May 2022)

• Record Type: Journal Article
• Authors: Bhunia, Rupam Kumar; Menard, Guillaume N.; Eastmond, Peter J.
• Is Part Of: FEBS letters. Volume 596:Issue 15(2022)
• Page Start: 1865
• Page End: 1870
• Publication Date: 2022-05-06
• Subjects: CRISPR/Cas9 -- genome editing -- promoter fusion -- transcriptional gain‐of‐function -- transgene‐free -- plant oil metabolism; Biochemistry -- Periodicals; Biophysics -- Periodicals; Molecular biology -- Periodicals; Biochimie -- Périodiques; Biochemistry; Biophysics; Molecular biology; Periodicals; 572.05
• DOI: 10.1002/1873-3468.14365
• Languages: English
• ISSNs: 0014-5793

Abstract

Abstract : 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. Abstract : Our study demonstrates a method to create a transcriptional gain‐of‐function without inserting foreign DNA into an organism. We bring a gene under the control of a promoter from an upstream gene by deleting the intervening genomic region using CRISPR/Cas9. We use this approach to engineer plant leaves to accumulate oil.