The DOI matches its Crossref record; 13 of the 13 references it cites were found in a registry or an index.
Identifying and Visualizing Functional PAM Diversity across CRISPR-Cas Systems
Open accessAbstract
SUMMARY CRISPR-Cas adaptive immune systems in prokaryotes boast a diversity of protein families and mechanisms of action, where most systems rely on protospacer-adjacent motifs (PAMs) for DNA target recognition. Here, we developed an in vivo, positive, and tunable screen termed PAM-SCANR (PAM screen achieved by NOT-gate repression) to elucidate functional PAMs as well as an interactive visualization scheme termed the PAM wheel to convey individual PAM sequences and their activities. PAM-SCANR and the PAM wheel identified known functional PAMs while revealing complex sequence-activity landscapes for the Bacillus halodurans I-C (Cascade), Escherichia coli I-E (Cascade), Streptococcus thermophilus II-A CRISPR1 (Cas9), and Francisella novicida V-A (Cpf1) systems. The PAM wheel was also readily applicable to existing high-throughput screens and garnered insights into SpyCas9 and SauCas9 PAM diversity. These tools offer powerful means of elucidating and visualizing functional PAMs toward accelerating our ability to understand and exploit the multitude of CRISPR-Cas systems in nature.
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Citations
Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
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BackgroundMethodInfluentialIn vivo genome editing using Staphylococcus aureus Cas9
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Cited by
CRISPR–Cas9 Structures and Mechanisms
Jiang, F., & Doudna, J. A. (2017) · Annual Review of Biophysics · doi:10.1146/annurev-biophys-062215-010822 · Cited by 2,029
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
Self-citationCRISPR-P 2.0: An Improved CRISPR-Cas9 Tool for Genome Editing in Plants
Liu, H., Ding, Y., Zhou, Y., Jin, W., Xie, K., & Chen, L.-L. (2017) · Molecular Plant · doi:10.1016/j.molp.2017.01.003 · Cited by 837
RNA-guided DNA insertion with CRISPR-associated transposases
Strecker, J., Ladha, A., Gardner, Z., Schmid-Burgk, J. L., Makarova, K. S., Koonin, E. V., & Zhang, F. (2019) · Science · doi:10.1126/science.aax9181 · Cited by 708
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Metadata
- Authors
- Leenay, R. T., Maksimchuk, K. R., Slotkowski, R. A., Agrawal, R. N., Gomaa, A. A., Briner, A. E., Barrangou, R., & Beisel, C. L.
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2016-04
- Volume
- 62
- Issue
- 1
- Pages
- 137-147
- ISSN
- 1097-2765 (ISSN-L)
- OpenAlex
- W2327182906
- Cited by
- 336
- References
- 13
- FWCI
- 18.96
- Type
- journal article
- Language
- en
- Open access
- bronze
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