Epigenetic features drastically impact CRISPR–Cas9 efficacy in plants

• DOI: 10.1093/plphys/kiac285
• Publication Date: 2022-06-11T13:33:54Z
• Authors: Trevor Weiss; Peter A Crisp; Krishan M Rai; Meredith Song; Nathan M Springer; Feng Zhang
• Keywords: Epigenomics; Gene Editing; 0301 basic medicine; 03 medical and health sciences; Regular Issue Content; Arabidopsis; CRISPR-Cas Systems; Chromatin

Abstract CRISPR–Cas9-mediated genome editing has been widely adopted for basic and applied biological research in eukaryotic systems. While many studies consider DNA sequences of CRISPR target sites as the primary determinant mutagenesis efficiency mutation profiles, increasing evidence reveals substantial role chromatin context. Nonetheless, most prior are limited by lack sufficient epigenetic resources and/or only transiently expressing CRISPR–Cas9 a short time window. In this study, we leveraged wealth high-resolution epigenomic Arabidopsis (Arabidopsis thaliana) to address impact features on using stable transgenic plants. Our results indicated that methylation could lead variations up 250-fold. Low efficiencies were mostly associated with repressive heterochromatic features. This effect appeared persist through cell divisions but be alleviated reduction at sites. Moreover, specific features, such H3K4me1, H3.3, H3.1, appear significant variation profiles mediated non-homologous end joining repair pathway. findings provide strong have lasting impacts both double-strand break outcomes.