Correction to: Fungal Biol Biotechnol (2019) 6:15 10.1186/s40694-019-0079-4
Following publication of the original article [1], the authors reported that Table 5 was missing in the published version, although it was originally submitted and reviewed along with the rest of the manuscript. The complete Table 5 is given in this erratum.
Table 5.
Practical guidance for the implementation of CRISPR technology in filamentous fungi based on data obtained for T. thermophilus in this study
Plasmid-based approach | RNP-based approach | |
---|---|---|
Preparation of nuclease | Cloning of the nuclease into a plasmid prior transformation is mandatory. When constitutively expressed, risk of off-targets should be considered. When present on AMA-plasmid, the risk should be lower but still present | Cloning of the nuclease into a plasmid allowing heterologous expression, e.g. in E. coli, is a prerequisite. Once established and purified, the nuclease can be aliquoted and stored prior to use. As the protein does not become expressed in the targeted fungus, the risk of off-targets should be very small |
Preparation of guide RNA | Plasmid-based, thus more stable during handling and storage | Involves in vitro transcription, hence potentially sensitive to handling errors |
Transformation procedure | Easy | Easy but requires preassembly of RNPs |
Transformation rate | Very high also with four targets |
Very high for single and double targets Low for three and four targets |
Single-targeting efficiency of FnCpf1, AsCpf1, SpCas9 | Locus-dependent | Locus-dependent |
Multiplex-targeting efficiency of FnCpf1 | High (34 ± 6% in this study) | Low (13 ± 2% in this study) |
MTP-based down-scaling for FnCpf1 | Possible with no loss in efficiency with respect to single and double targeting | Possible with no loss in efficiency with respect to single targetinga |
aDouble targeting was not tested
The original article has been corrected.
Footnotes
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Reference
- 1.Kwon MJ, Schütze T, Spohner S, Haefner S, Meyer V. Practical guidance for the implementation of the CRISPR genome editing tool in filamentous fungi. Fungal Biol Biotechnol. 2019;6:15. doi: 10.1186/s40694-019-0079-4. [DOI] [PMC free article] [PubMed] [Google Scholar]