<i>In-planta</i> Gene Targeting in Barley Using Cas9 With and Without Geminiviral Replicons.

ABSTRACT: Advances in the use of RNA-guided Cas9-based genome editing in plants have been rapid over the last few years. A desirable application of genome editing is gene targeting (GT), as it allows a wide range of precise modifications; however, this remains inefficient especially in key crop species. Here, we describe successful, heritable gene targeting in barley at the target site of Cas9 using an in-planta strategy but fail to achieve the same using a wheat dwarf virus replicon to increase the copy number of the repair template. Without the replicon, we were able to delete 150 bp of the coding sequence of our target gene whilst simultaneously fusing in-frame mCherry in its place. Starting from 14 original transgenic plants, two plants appeared to have the required gene targeting event.

SUBMITTER: Lawrenson T 

PROVIDER: S-EPMC8525372 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature