Multiplexed CRISPR-Cas9 mutagenesis of rice <i>PSBS1</i> noncoding sequences for transgene-free overexpression.

ABSTRACT: Understanding CRISPR-Cas9's capacity to produce native overexpression (OX) alleles would accelerate agronomic gains achievable by gene editing. To generate OX alleles with increased RNA and protein abundance, we leveraged multiplexed CRISPR-Cas9 mutagenesis of noncoding sequences upstream of the rice PSBS1 gene. We isolated 120 gene-edited alleles with varying non-photochemical quenching (NPQ) capacity in vivo-from knockout to overexpression-using a high-throughput screening pipeline. Overexpression increased OsPsbS1 protein abundance two- to threefold, matching fold changes obtained by transgenesis. Increased PsbS protein abundance enhanced NPQ capacity and water-use efficiency. Across our resolved genetic variation, we identify the role of 5'UTR indels and inversions in driving kn

SUBMITTER: Patel-Tupper D 

PROVIDER: S-EPMC11160471 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature