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Marker-free carotenoid-enriched rice generated through targeted gene insertion using CRISPR-Cas9.

Abstract：

:Targeted insertion of transgenes at pre-determined plant genomic safe harbors provides a desirable alternative to insertions at random sites achieved through conventional methods. Most existing cases of targeted gene insertion in plants have either relied on the presence of a selectable marker gene in the insertion cassette or occurred at low frequency with relatively small DNA fragments (<1.8 kb). Here, we report the use of an optimized CRISPR-Cas9-based method to achieve the targeted insertion of a 5.2 kb carotenoid biosynthesis cassette at two genomic safe harbors in rice. We obtain marker-free rice plants with high carotenoid content in the seeds and no detectable penalty in morphology or yield. Whole-genome sequencing reveals the absence of off-target mutations by Cas9 in the engineered plants. These results demonstrate targeted gene insertion of marker-free DNA in rice using CRISPR-Cas9 genome editing, and offer a promising strategy for genetic improvement of rice and other crops.

journal_name

Nat Commun

journal_title

Nature communications

authors

Dong OX,Yu S,Jain R,Zhang N,Duong PQ,Butler C,Li Y,Lipzen A,Martin JA,Barry KW,Schmutz J,Tian L,Ronald PC

doi

10.1038/s41467-020-14981-y

subject

Has Abstract

pub_date

2020-03-04 00:00:00

pages

1178

issue

1

issn

2041-1723

pii

10.1038/s41467-020-14981-y

journal_volume

11

pub_type

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