Leveraging CRISPR Cas9 RNPs and Cre-<i>loxP</i> in <i>Picochlorum celeri</i> for generation of field deployable strains and selection marker recycling.

ABSTRACT: As new highly productive strains of algae are discovered and developed to meet the energy, chemical, and food requirements of the future, genetic engineering of those strains in a manner that yields deployable transformants is paramount. This study introduces the novel CRoxP (Cas9 RNPs coupled with an inducible CRe-loxP) system for rapid generation of marker- and transgene-free strains of Picochlorum celeri. The CRoxP system allows reuse of selection markers without Cas9 expression in vivo, eliminating many of the bottlenecks associated with conventional CRISPR Cas9 use for precise genome editing. In P. celeri, transformants were generated with a turnaround time as short as 21 days between transformation and being ready for another round of transformation with the same

SUBMITTER: Burch TA 

PROVIDER: S-EPMC12301381 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature