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Mode of Action (MoA)
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binds to sgRNA loaded Cas9 preventing the complex from binding target DNA
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Evidence
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AcrIIC6Nme was identified using a guilt-by-association bioinformatic approach and tested for anti-CRISPR activity via in vivo phage targeting assays. Expression of AcrIIC6Nme in E. coli co-expressing Nme1Cas9 and an sgRNA targeting phage Mu restored phage infectivity, indicating inhibition of CRISPR interference. This inhibition was selective, effective only against closely related type II-C Cas9s (Nme1Cas9, Nme2Cas9, HpaCas9, BoeCas9), but not distantly related variants (GeoCas9, CjeCas9, CdiCas9). In vivo pulldown (ECO_0006249) assays demonstrated that AcrIIC6Nme selectively co-purifies with the Nme1Cas9:sgRNA complex but not apo-Cas9, indicating specific binding to the Cas9:sgRNA binary complex. These results support the activity of AcrIIC6Nme as a selective type II-C anti-CRISPR that binds the loaded Cas9 complex and inhibits DNA targeting.
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MoA Category
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binds and inhibits host defence system
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Subtype(s) of the defence system(s) inhibited by the protein
Defence Subtype
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Neisseria meningitidis type II-C CRISPR-Cas
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Relevant publication(s)
DOI
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No DOI, master thesis (Khan, A.N., 2021. Characterizing a Novel Type II-C Anti-CRISPR, AcrIIC6)
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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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-
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Genome(s) encoding the protein
Protein Source
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Prophages in Neisseria meningitidis and Pasteurella multocida
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Defence system(s) inhibited by the protein
Defences
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CRISPR-Cas
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