Expression analysis of red clover roots in CRISPRCas9 C2H2 ZFP knockout lines following rhizobium inoculation

Red clover knockout lines were recovered following transformation using CRISPR/Cas9 vectors targeting the LOC123890030, LOC123902409 and LOC123884023 genes. Homozygous (LOC123884023) and hemizygous (LOC123890030 and LOC123902409) knockout plants were recovered. Cuttings from the transgenic clones were rooted and inoculated with Rhizobium leguminosarum biovar trifolii, (strain USDA 2046). RNA was isolated from roots three weeks post-inoculation from both inoculated and uninoculated plants to compared gene expression of the nodulating plants by RNA-seq.