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

A - Papers appearing in refereed journals

Bhunia, R. K., Menard, G. N. and Eastmond, P. J. 2022. 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. https://doi.org/10.1002/1873-3468.14365

AuthorsBhunia, R. K., Menard, G. N. and Eastmond, P. J.
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 non-essential 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 twenty-fold and the triacylglycerol content increases by around thirty-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.

KeywordsTransgene-free; CRISPR/Cas9; Genome editing; Genomic deletion; Promoter fusion; Transcriptional gain-of-function; Metabolic engineering; Plant oil metabolism
Year of Publication2022
JournalFEBS Letters
Digital Object Identifier (DOI)https://doi.org/10.1002/1873-3468.14365
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeTailoring Plant Metabolism ISP
Publisher's version
Accepted author manuscript
Supplemental file
Supplemental file
Output statusPublished
Publication dates
Online01 May 2022
Publication process dates
Accepted19 Apr 2022
PublisherFEBS Press
ISSN0014-5793

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