The DOI matches its Crossref record; 25 of the 25 references it cites were found in a registry or an index.
A gRNA-tRNA array for CRISPR-Cas9 based rapid multiplexed genome editing in Saccharomyces cerevisiae
Open accessAbstract
With rapid progress in DNA synthesis and sequencing, strain engineering starts to be the rate-limiting step in synthetic biology. Here, we report a gRNA-tRNA array for CRISPR-Cas9 (GTR-CRISPR) for multiplexed engineering of Saccharomyces cerevisiae. Using reported gRNAs shown to be effective, this system enables simultaneous disruption of 8 genes with 87% efficiency. We further report an accelerated Lightning GTR-CRISPR that avoids the cloning step in Escherichia coli by directly transforming the Golden Gate reaction mix to yeast. This approach enables disruption of 6 genes in 3 days with 60% efficiency using reported gRNAs and 23% using un-optimized gRNAs. Moreover, we applied the Lightning GTR-CRISPR to simplify yeast lipid networks, resulting in a 30-fold increase in free fatty acid production in 10 days using just two-round deletions of eight previously identified genes. The GTR-CRISPR should be an invaluable addition to the toolbox of synthetic biology and automation.
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Cites
DNA targeting specificity of RNA-guided Cas9 nucleases
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Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system
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The CRISPR/Cas9 system for plant genome editing and beyond
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Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs
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Multiplex metabolic pathway engineering using CRISPR/Cas9 in Saccharomyces cerevisiae
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Selection of chromosomal DNA libraries using a multiplex CRISPR system
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Past, present, and future of CRISPR genome editing technologies
Pacesa, M., Pelea, O., & Jinek, M. (2024) · Cell · doi:10.1016/j.cell.2024.01.042 · Cited by 609
Zebrafish patient avatars in cancer biology and precision cancer therapy
Fazio, M., Ablain, J., Chuan, Y., Langenau, D. M., & Zon, L. I. (2020) · Nature Reviews Cancer · doi:10.1038/s41568-020-0252-3 · Cited by 257
Direct-seq: programmed gRNA scaffold for streamlined scRNA-seq in CRISPR screen
Song, Q., Ni, K., Liu, M., Li, Y., Wang, L., Wang, Y., Liu, Y., Yu, Z., Qi, Y., Lu, Z., et al. (2020) · Genome Biology · doi:10.1186/s13059-020-02044-w · Cited by 26
Metadata
- Authors
- Zhang, Y., Wang, J., Wang, Z., Zhang, Y., Shi, S., Nielsen, J., & Liu, Z.
- Venue
- Nature Communications
- Published
- 2019-03-05
- Volume
- 10
- Issue
- 1
- Pages
- 1053-1053
- ISSN
- 2041-1723 (ISSN-L)
- OpenAlex
- W2920169518
- Cited by
- 294
- References
- 25
- FWCI
- 10.24
- Type
- journal article
- Language
- en
- Open access
- gold
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- Zhang, Y., Wang, J., Wang, Z., Zhang, Y., Shi, S., Nielsen, J., & Liu, Z. (2019). A gRNA-tRNA array for CRISPR-Cas9 based rapid multiplexed genome editing in Saccharomyces cerevisiae. Nature Communications. https://doi.org/10.1038/s41467-019-09005-3
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