The DOI matches its Crossref record; 8 of the 8 references it cites were found in a registry or an index.
CRISPR–Cas9-assisted recombineering in Lactobacillus reuteri
Open accessAbstract
Clustered regularly interspaced palindromic repeats (CRISPRs) and the CRISPR-associated (Cas) nuclease protect bacteria and archeae from foreign DNA by site-specific cleavage of incoming DNA. Type-II CRISPR-Cas systems, such as the Streptococcus pyogenes CRISPR-Cas9 system, can be adapted such that Cas9 can be guided to a user-defined site in the chromosome to introduce double-stranded breaks. Here we have developed and optimized CRISPR-Cas9 function in the lactic acid bacterium Lactobacillus reuteri ATCC PTA 6475. We established proof-of-concept showing that CRISPR-Cas9 selection combined with single-stranded DNA (ssDNA) recombineering is a realistic approach to identify at high efficiencies edited cells in a lactic acid bacterium. We show for three independent targets that subtle changes in the bacterial genome can be recovered at efficiencies ranging from 90 to 100%. By combining CRISPR-Cas9 and recombineering, we successfully applied codon saturation mutagenesis in the L. reuteri chromosome. Also, CRISPR-Cas9 selection is critical to identify low-efficiency events such as oligonucleotide-mediated chromosome deletions. This also means that CRISPR-Cas9 selection will allow identification of recombinant cells in bacteria with low recombineering efficiencies, eliminating the need for ssDNA recombineering optimization procedures. We envision that CRISPR-Cas genome editing has the potential to change the landscape of genome editing in lactic acid bacteria, and other Gram-positive bacteria.
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Cites
Multiplex Genome Engineering Using CRISPR/Cas Systems
Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A., et al. (2013) · Science · doi:10.1126/science.1231143 · Cited by 15,339
CRISPR-Cas systems for editing, regulating and targeting genomes
Sander, J. D., & Joung, J. K. (2014) · Nature Biotechnology · doi:10.1038/nbt.2842 · Cited by 3,153
RNA-guided editing of bacterial genomes using CRISPR-Cas systems
Jiang, W., Bikard, D., Cox, D., Zhang, F., & Marraffini, L. A. (2013) · Nature Biotechnology · doi:10.1038/nbt.2508 · Cited by 2,429
Improving CRISPR-Cas nuclease specificity using truncated guide RNAs
Fu, Y., Sander, J. D., Reyon, D., Cascio, V. M., & Joung, J. K. (2014) · Nature Biotechnology · doi:10.1038/nbt.2808 · Cited by 1,993 · cited 1×
“pyogenes CRISPR–Cas9 system cleaves target DNA efficiently if non-homologous bases at position 21–28 are present in the protospacer, whereas a protospacer of 17 base pairs still yields efficient targeting by Cas9 (43,44).”
BackgroundRNA-guided genetic silencing systems in bacteria and archaea
Wiedenheft, B., Sternberg, S. H., & Doudna, J. A. (2012) · Nature · doi:10.1038/nature10886 · Cited by 1,851
Programming cells by multiplex genome engineering and accelerated evolution
Wang, H. H., Isaacs, F. J., Carr, P. A., Sun, Z. Z., Xu, G., Forest, C. R., & Church, G. M. (2009) · Nature · doi:10.1038/nature08187 · Cited by 1,529 · cited 1×
“Also, our work combined with current developments in CRISPR–Cas9 genome editing makes the application of MAGE in lactobacilli and bifidobacteria tantalizing, yet not unreasonable (45,46).”
BackgroundProbiotic L. reuteri Treatment Prevents Bone Loss in a Menopausal Ovariectomized Mouse Model
Britton, R. A., Irwin, R., Quach, D., Schaefer, L., Zhang, J., Lee, T., Parameswaran, N., & McCabe, L. R. (2014) · Journal of Cellular Physiology · doi:10.1002/jcp.24636 · Cited by 499
Thomas, C. M., Hong, T., van Pijkeren, J. P., Hemarajata, P., Trinh, D. V., Hu, W., Britton, R. A., Kalkum, M., & Versalovic, J. (2012) · PLoS ONE · doi:10.1371/journal.pone.0031951 · Cited by 491
Cited by
Applications of CRISPR technologies in research and beyond
Barrangou, R., & Doudna, J. A. (2016) · Nature Biotechnology · doi:10.1038/nbt.3659 · Cited by 1,005
CRISPR-Cas immunity in prokaryotes
Marraffini, L. A. (2015) · Nature · doi:10.1038/nature15386 · Cited by 829
Ishino, Y., Krupovic, M., & Forterre, P. (2018) · Journal of Bacteriology · doi:10.1128/jb.00580-17 · Cited by 558
Sun, Z., Harris, H. M. B., McCann, A., Guo, C., Argimón, S., Zhang, W., Yang, X., Jeffery, I. B., Cooney, J. C., Kagawa, T. F., et al. (2015) · Nature Communications · doi:10.1038/ncomms9322 · Cited by 543
Microbiome and Human Health: Current Understanding, Engineering, and Enabling Technologies
Aggarwal, N., Kitano, S., Puah, G. R. Y., Kittelmann, S., Hwang, I. Y., & Chang, M. W. (2023) · Chemical Reviews · doi:10.1021/acs.chemrev.2c00431 · Cited by 508
Metadata
- Authors
- Oh, J.-H., & van Pijkeren, J.-P.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2014-07-29
- Volume
- 42
- Issue
- 17
- Pages
- e131-e131
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2119633529
- Cited by
- 392
- References
- 8
- FWCI
- 10.95
- Type
- journal article
- Language
- en
- Open access
- gold
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