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Mode of Action (MoA)
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sequesters cyclic dinucleotides and gcADPR.
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Evidence
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A synthetic E. coli strain was engineered with a genetic circuit where SpCas9 targeted a plasmid-borne chloramphenicol resistance gene. If an anti-CRISPR protein inhibits Cas9, the strain retains chloramphenicol resistance and survives antibiotic selection (ECO_0007003). In vitro cleavage assay evidence (a biochemical assay where purified Cas9 protein and guide RNA are incubated with target DNA and a candidate ACR protein outside of any living cell) further supported anti-CRISPR activity. However, direct binding is not supported by biolayer Interferometry evidence (ECO_0006350). Another study was not able to reproduce anti-CRISPR activity (10.1038/s41586-024-08122-4). Gel Shift Assay (ECO_0000096) was used to test binding to cyclic oligonucleotides, affinity was measured by Surface Plasmon Resonance (ECO_0001127), and there is X-ray crystallography evidence (ECO_0005670) to support the binding.
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MoA Category
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degrades or sequesters molecules utilised by host defence systems
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Subtype(s) of the defence system(s) inhibited by the protein
Defence Subtype
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Pseudomonas aeruginosa strain PAO1 type I (ThsA with SIR2) Thoeris
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Relevant publication(s)
DOI
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10.1016/j.chom.2019.01.003,
10.1038/s41586-024-08122-4
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Other components of the anti-defence system
Multicomponent System
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-
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Known structure in PDB
PDB ID
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8KBI
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Genome(s) encoding the protein
Protein Source
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Metagenome
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Defence system(s) inhibited by the protein
Defences
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CBASS,
Thoeris
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