Methods in molecular biology (Clifton, N.J.)Oct 1, 2025

Using CRISPR to Edit Pichia pastoris Strains

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Oct 2, 2025

Abstract

The CRISPR/Cas9 system is a popular tool for editing genes and improving how organisms function. In this paper, we share a straightforward method for using CRISPR to make precise changes in the genome of Pichia pastoris, a type of yeast used in industry. Our protocol shows very high success in disabling specific genes, which can help scientists create better strains for various applications.

Simplified

The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats, CRISPR-associated protein 9) system has become a commonly used tool for genome editing and metabolic engineering. For Komagataella phaffii, commercialized as Pichia pastoris, the CRISPR/Cas9 protocol for genome editing was established in 2016 and since then has been employed to facilitate genetic modifications such as markerless gene disruptions and deletions as well as to enhance the efficiency of homologous recombination.In this chapter, we describe a robust basic protocol for CRISPR-based genome editing, demonstrating near 100% targeting efficiency for gene inactivation via a frameshift mutation. As described in other chapters of this volume, CRISPR/Cas9 technologies for use in P. pastoris have been further optimized for various specific purposes.

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Using CRISPR to Edit Pichia pastoris Strains

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You found one interesting study. We’ll send the next 7.

What Lands in Your Inbox Each Week:

Recent issues from the crispr gene editing brief