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Updated: Aug 30, 2025

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis
Published on: June 9, 2020
Genome editing in Penicillium funiculosum using in vitro assembled CRISPR-Cas9 ribonucleoprotein complexes
Tulika Sinha1, Syed Shams Yazdani2
1Microbial Engineering Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India.
Abstract:
The plasmid-free CRISPR-Cas9-based genome editing in fungi is a precise and time-saving approach. Here, we present a detailed protocol for genetic manipulation in Penicillium funiculosum, which includes design and synthesis of sgRNA, high-quality protoplast preparation, and PEG-mediated protoplast transformation of linear donor DNA along with in vitro synthesized RNP complex composed of sgRNA and host-specific Cas9. This technique is beneficial for researchers interested in functional analysis of genes as it improves reproducibility and replicability of the experiment. For complete details on the use and execution of this protocol, please refer to Randhawa et al. (2021).
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