CRISPR-Cas9-Mediated Mutagenesis of the Rubisco Small Subunit Family in <i>Nicotiana tabacum</i>.

ABSTRACT: Engineering the small subunit of the key CO₂-fixing enzyme Rubisco (SSU, encoded by rbcS) in plants currently poses a significant challenge, as many plants have polyploid genomes and SSUs are encoded by large multigene families. Here, we used CRISPR-Cas9-mediated genome editing approach to simultaneously knock-out multiple rbcS homologs in the model tetraploid crop tobacco (Nicotiana tabacum cv. Petit Havana). The three rbcS homologs rbcS_S1a, rbcS_S1b and rbcS_T1 account for at least 80% of total rbcS expression in tobacco. In this study, two multiplexing guide RNAs (gRNAs) were designed to target homologous regions in these three genes. We generated tobacco mutant lines with indel mutations in all three genes, including one line

SUBMITTER: Donovan S 

PROVIDER: S-EPMC8525408 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature