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A versatile anti-CRISPR platform for opto- and chemogenetic control of CRISPR-Cas9 and Cas12 across a wide range of orthologs.

Brenker L, Aschenbrenner S, Bubeck F, Staykov K, Gebhardt C, Wolf B, Jendrusch M, Kroll AS, Becker J, Ambiel I, Fackler OT, Grimm D, Mathony J, Niopek D
Nucleic Acids Res. 2025 Jul 19;53(14):gkaf752. doi: 10.1093/nar/gkaf752. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
246043CMV_AcrIIA5_AsLOV2-D41Expression of AcrIIA5-AsLOV2 hybrid in mammalian cells; insertion before D41; enables optogenetic control of SauCas9
246044CMV_AcrIIA5_AsLOV2-N77Expression of AcrIIA5-AsLOV2 hybrid in mammalian cells; insertion before N77; enables optogenetic control of SauCas9
246045CMV_AcrIIA5_cpGR2(GGS5)-D41Expression of AcrIIA5-cpGR2 hybrid in mammalian cells; insertion before D41; enables chemogenetic control of SauCas9
246046CMV_AcrIIA5_cpGR2(GGS5)-N77Expression of AcrIIA5-cpGR2 hybrid in mammalian cells; insertion before N77; enables chemogenetic control of SauCas9
246048AAV_CMV-AcrIIA5_cpGR2(GGS5)-N77Expression of AcrIIA5-cpGR2 hybrid in mammalian cells; insertion before N77; enables chemogenetic control of SauCas9; contains ITRs for AAV packaging with Rep2
246049AcrVA1_AsLOV2-S103ΔELSExpression of AcrVA1-AsLOV2 hybrid in mammalian cells; insertion before S103 with deletion of surrounding amino acids ELS; enables optogenetic control of MbCas12a
246050AcrVA1_cpGR2(GGS5)-S103ΔELSExpression of AcrVA1-cpGR2 hybrid in mammalian cells; insertion before S103 with deletion of surrounding amino acids ELS; enables chemogenetic control of MbCas12a
246051AcrVA1_cpGR2(GGS5)-S103ΔELS_(GGS)2-(GGS)2Expression of AcrVA1-cpGR2 hybrid in mammalian cells; insertion before S103 with deletion of surrounding amino acids ELS and GGS2 linkers; enables chemogenetic control of MbCas12a
246052CMV_AcrIIA4_cpGR2(GGS5)-ΔN64_Q65_E66Expression of AcrIIA4-cpGR2 hybrid in mammalian cells after Bubeck et al Nat Methods 2018; enables chemogenetic control of SpyCas9
246053CMV_AcrIIC3_cpGR2(GGS5)-F59Expression of AcrIIC3-cpGR2 hybrid in mammalian cells; insertion before F59; enables chemogenetic control of NmeCas9
246054H1_sgRNA(GRIN2B)_SauCas9Expression of a sgRNA targeting the GRIN2B locus and SauCas9 under the same pol-III H1 promoter
246055H1_sgRNA(GRIN2B)_SauCas9_AcrIIA5-AsLOV2(D41)Expression of a sgRNA targeting the GRIN2B locus, a SauCas9 and an AcrIIA5-cpGR2 hybrid (insertion before D41) linked through a P2A sequence, under the same pol-III H1 promoter
246056H1_sgRNA(GRIN2B)_SauCas9_AcrIIA5-AsLOV2(N77)Expression of a sgRNA targeting the GRIN2B locus, a SauCas9 and an AcrIIA5-cpGR2 hybrid (insertion before N77) linked through a P2A sequence, under the same pol-III H1 promoter

Antibodies from Article