Journal Article

CRISPR/Cas9 induces exon skipping that facilitates development of fragrant rice

Tang, Yongchao, Li, Jinbo, Wang, Fujun, Wang, Chunlian, Abdelrahman, Mohamed, Zhao, Kaijun, Ji, Zhiyuan, Qi, Huaxiong

2021

Overview

[...]our results showed that exon skipping occurred due to the CRISPR/Cas9‐mediated alterations at the splice site of a plant gene. [...]we found the exon 2 removal during the splicing process in the Rbadh2ΔG and Rbadh2ΔAAG mutants caused shifting of reading frame in the processed mRNA, resulting in a PTC in the exon 3 (Figure 1g). The transcripts containing a PTC will be degraded by non‐sense‐mediated decay (Capito et al., 2018), as indicated by the reduced expression level of the mutated Osbadh2 compared with the WT (Figure 1d, h). Since loss of OsBADH2 function promotes accumulation of 2AP, we explored the consequence of Osbadh2 exon 2 skipping on the 2AP content in grains of the homozygous transgene‐free plants of Rbadh2ΔG and Rbadh2ΔAAG using gas chromatography–mass spectrometry (GC‐MS). [...]this study provides the first evidence that CRISPR/Cas9‐mediated deletion of the exonic nucleotide at the exon–intron junction of a plant gene could cause exon skipping during pre‐mRNA splicing.

• Publisher: John Wiley & Sons, Inc,John Wiley and Sons Inc
• Subject: alternative splicing / BADH2 / Brief Communication / Brief Communications / Cas9 / Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR / CRISPR-Cas Systems - genetics / Exon skipping / Exons - genetics / fragrant rice / Gas chromatography / Gene deletion / Gene expression / Mass spectrometry / Mass spectroscopy / mis‐splicing / Mutation / Nucleotides / Oryza - genetics / premature termination codon / Retention / Rice / Splicing / Standard deviation / Transgenic plants