CRISPR/Cas Genome Editing in Filamentous Fungi

Current advancement in the application of prime editing

“…In the same year, the genome of Streptococcus pneumoniae and E. coli were successfully edited by using the CRISPR-Cas9 system ( Jiang et al, 2013 ). Subsequently, the CRISPR-Cas9 system has been applied in a variety of filamentous fungi ( Song et al, 2019 ; Rozhkova and Kislitsin, 2021 ). In 2015, the CRISPR-Cas system was applied in Aspergillus nidulans ( Nødvig et al, 2015 ), following application in Penicillium chrysogenum , Trichoderma reesei , Neurospora crassa , Acremonium chrysogenum and so on ( Liu et al, 2015 ; Matsu-Ura et al, 2015 ; Pohl et al, 2016 ; Chen and Chu, 2019 ; Chen et al, 2020 ).…”

The expanding CRISPR toolbox for natural product discovery and engineering in filamentous fungi

“…59,60 Several genetic parts have been used for in fungus expression of Cas9 and Cas12a across fungal taxa, such as the promoters Ptef1, PgpdA, and PtrpC, PtetON. [10][11][12]61 In some reports the cas gene is codon optimised, while human codon-optimised cas9 and cas12a genes have also been shown to be functional in some lamentous fungi. 10,61 Importantly, some considerations need to be taken for the correct expression of gRNAs in vivo.…”

“…18 Amongst the diverse variety of naturally evolved CRISPR/Cas systems, the type II systems Cas9 and Cas12a have been adapted for gene editing in filamentous fungi. 11,12 The important components shared by these systems are an RNA-guided Cas nuclease that can be directed to a DNA target specified by base-pairing complementarity with a CRISPR RNA (crRNA). 19 Each crRNA is composed of a scaffold region and a ∼20 nt spacer sequence that can be programmed to define the DNA target.…”

“…The development and general applications of CRISPR/Cas systems in lamentous fungi have been covered elsewhere. [8][9][10][11][12] In this review, we highlight the innovative approaches by which CRISPR/Cas tools can be used for SM genome mining and engineering. Firstly, we will provide a bird's-eye view of the principles of CRISPR/Cas experimental design, execution and analysis used across BGC manipulation reports.…”

Evaluation of Aspergillus niger Six Constitutive Strong Promoters by Fluorescent-Auxotrophic Selection Coupled with Flow Cytometry: A Case for Citric Acid Production

“…With the rapid development of systems biology and synthetic biology, rational engineering has gradually become the dominant approach to reconstruct A. niger cell factories with higher titer, yield, and productivity [ 2 , 4 , 5 ]. Especially the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system-mediated genome-editing techniques [ 6 , 7 , 8 ] have broken the bottleneck of highly efficient genetic manipulation.…”