Cas12a RNP-mediated co-transformation enables transgene-free multiplex genome editing, long deletions, and inversions in citrus chromosome.

ABSTRACT:

Introduction

Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), is a devastating disease worldwide. Previously, we successfully generated canker-resistant Citrus sinensis cv. Hamlin lines in the T0 generation. This was achieved through the transformation of embryogenic protoplasts using the ribonucleoprotein (RNP) containing Cas12a and one crRNA to edit the canker susceptibility gene, CsLOB1, which led to small indels.

Methods

Here, we transformed embryogenic protoplasts of Hamlin with RNP containing Cas12a and three crRNAs.

Results

Among the 10 transgene-free genome-edited lines, long deletions were obtained in five lines. Additionally, inversions were observed in three of the five edited lines with long deletions, but not in a

SUBMITTER: Su H 

PROVIDER: S-EPMC11324552 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature