ArticleMicrobial cell factories2018
Gene repression via multiplex gRNA strategy in Y. lipolytica.
Article in Microbial cell factories, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed.
- 10 years of CRISPR/CAS genomic engineering in Yarrowia lipolytica.Bioprocess and biosystems engineering · 2026Review
- Transcriptional regulation: efficient genetic engineering tools for non-conventional yeasts.FEMS yeast research · 2026Review
- Unlocking the Synthetic Potential of Yarrowia lipolytica: Innovating Gene Expression Tools.Microbial biotechnology · 2025Review
- Establishment of CRISPR-STAR System to Realise Simultaneous Transcriptional Activation and Repression in Yarrowia lipolytica.Microbial biotechnology · 2025Article
- Reprogramming yeast metabolism for customized starch-rich micro-grain through low-carbon microbial manufacturing.Nature communications · 2025Article
- Advances and perspectives in genetic expression and operation for the oleaginous yeastSynthetic and systems biotechnology · 2024Review
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- ReprogrammingScience advances · 2024Article
- Blunted blades: new CRISPR-derived technologies to dissect microbial multi-drug resistance and biofilm formation.mSphere · 2024Review
- CRISPR-based screening of small RNA modulators of bile susceptibility inProceedings of the National Academy of Sciences of the United States of America · 2024Article
- Genome-scale CRISPRi screening: A powerful tool in engineering microbiology.Engineering microbiology · 2023Review
- Recent Advances in CRISPR-Cas Technologies for Synthetic Biology.Journal of microbiology (Seoul, Korea) · 2023Review
- Quantitative and modularized CRISPR/dCas9-dCpf1 dual function system inFrontiers in bioengineering and biotechnology · 2023Article
- Development and application of CRISPR-based genetic tools in Bacillus species and Bacillus phages.Journal of applied microbiology · 2022Review
- Development of a gRNA Expression and Processing Platform for Efficient CRISPR-Cas9-Based Gene Editing and Gene Silencing in Candida tropicalis.Microbiology spectrum · 2022Article
- SynBioStrainFinder: A microbial strain database of manually curated CRISPR/Cas genetic manipulation system information for biomanufacturing.Microbial cell factories · 2022Article
- CRISPRi enables fast growth followed by stable aerobic pyruvate formation inEngineering in life sciences · 2022Article
- Non-homologous End Joining-Mediated Insertional Mutagenesis Reveals a Novel Target for Enhancing Fatty Alcohols Production inFrontiers in microbiology · 2022Article
- Recent Molecular Tools for the Genetic Manipulation of Highly Industrially Important Mucoromycota Fungi.Journal of fungi (Basel, Switzerland) · 2021Review
- Establishment of genomic library technology mediated by non-homologous end joining mechanism in Yarrowia lipolytica.Science China. Life sciences · 2021Article
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8 authors.
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Abstract
backgroundThe oleaginous yeast Yarrowia lipolytica is a promising microbial cell factory due to their biochemical characteristics and native capacity to accumulate lipid-based chemicals. To create heterogenous biosynthesis pathway and manipulate metabolic flux in Y. lipolytica, numerous studies have been done for developing synthetic biology tools for gene regulation. CRISPR interference (CRISPRi), as an emerging technology, has been applied for specifically repressing genes of interest.
resultsIn this study, we established CRISPRi systems in Y. lipolytica based on four different repressors, that was DNase-deactivated Cpf1 (dCpf1) from Francisella novicida, deactivated Cas9 (dCas9) from Streptococcus pyogenes, and two fusion proteins (dCpf1-KRAB and dCas9-KRAB). Ten gRNAs that bound to different regions of gfp gene were designed and the results indicated that there was no clear correlation between the repression efficiency and targeting sites no matter which repressor protein was used. In order to rapidly yield strong gene repression, a multiplex gRNAs strategy based on one-step Golden-brick assembly technology was developed. High repression efficiency 85% (dCpf1) and 92% (dCas9) were achieved in a short time by making three different gRNAs towards gfp gene simultaneously, which avoided the need of screening effective gRNA loci in advance. Moreover, two genes interference including gfp and vioE and three genes repression including vioA, vioB and vioE in protodeoxy-violaceinic acid pathway were also realized.
conclusionTaken together, successful CRISPRi-mediated regulation of gene expression via four different repressors dCpf1, dCas9, dCpf1-KRAB and dCas9-KRAB in Y. lipolytica is achieved. And we demonstrate a multiplexed gRNA targeting strategy can efficiently achieve transcriptional simultaneous repression of several targeted genes and different sites of one gene using the one-step Golden-brick assembly. This timesaving method promised to be a potent transformative tool valuable for metabolic engineering, synthetic biology, and functional genomic studies of Y. lipolytica.
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