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CRISPR Plasmids: RNA Targeting

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CRISPR Resources

Type VI CRISPR systems, including the enzymes Cas13a/C2c2 and Cas13b, target RNA rather than DNA. In bacteria, once they have recognized and cleaved the target RNA sequence, they adopt an enzymatically active state and can bind and cleave additional RNAs regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which require that each DNA target have high sequence identity to the spacer sequence and contain a PAM sequence just downstream of the sequence to be cleaved. This non-specific cleavage is thought to activate programmed cell death or dormancy for phage-infected bacterial cells so as to limit the spread of infection throughout the entire population.

Type VI enzymes that function in mammalian cells can be used to attentuate RNA levels. In mammalian systems, Cas13a does not exhibit the collateral RNA degradation seen in bacteria.

CRISPR RNA targeting schematic


Mammalian

Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
pC014 - LwCas13a-msfGFPLwCas13a-msfGFP (Homo sapiens)EF1a Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses active LwCas13a for mammalian RNA targeting lenti(AMP)
pC015 - dLwCas13a-NFdLwCas13a-eGFP-ZF-KRAB (Homo sapiens)CAG Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses negative-feedback dLwCas13a for mammalian RNA targeting/imaging pCAGEN
pC016 - LwCas13a guide expression backbone with U6 promoterLwCas13a Guide site (Synthetic) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Backbone for cloning LwCas13a guides under U6 promoter pUC19
pC017 - LwCas13a guide expression backbone with tRNAval promoterLwCas13a Guide site (Synthetic) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Backbone for cloning LwCas13a guides under tRNAval promoter pUC19
pC034 - LwCas13a-msfGFP-2A-BlastLwCas13a-msfGFP (Homo sapiens)EFSBlasticidin Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses active LwCas13a for mammalian RNA targeting with Blasticidin selection marker pFUGW
pC0040-LwaCas13a crRNA backboneU6-LwaCas13aDR-BbsI-BbsI-polyTU6 Zhang RNA editing with CRISPR-Cas13. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. Backbone for cloning guides that are compatible with LwaCas13a. Contains a 5' direct repeat. Clone using BbsI (BpiI). F overhang AAAC. R overhang AAAA pUC19
pC0043-PspCas13b crRNA backboneU6-PspCas13b DR-BbsI-BbsI-polyT Zhang RNA editing with CRISPR-Cas13. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. For cloning of guide RNAs compatible with PspCas13b. Contains a 3' direct repeat. Clone using BbsI (BpiI). F overhang cacc. R overhang caac pUC19
pC0044-EF1a-RanCas13b-NES-mapkRanCas13b (Other) Zhang RNA editing with CRISPR-Cas13. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. Expresses RanCas13b in mammalian cells for knockdown of target RNAs in combination with compatible guide RNAs. Localized to the cytoplasm EF1a-lenti
pC0045-EF1a-PguCas13b-NES-HIVPguCas13b (Other) Zhang RNA editing with CRISPR-Cas13. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. Expresses PguCas13b in mammalian cells for knockdown of target RNAs in combination with compatible guide RNAs. Localized to the cytoplasm EF1a-lenti
pC0046-EF1a-PspCas13b-NES-HIVPspCas13b (Other) Zhang RNA editing with CRISPR-Cas13. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. Expresses PspCas13b in mammalian cells for knockdown of target RNAs in combination with compatible guide RNAs. Localized to the cytoplasm EF1a-lenti
pC0049-EF1a-dPSPCas13b-NES-HIV, H133A/H1058APspCas13b (Other)EF1a Zhang RNA editing with CRISPR-Cas13. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. Catalytically inactive PSPCas13b. RNase activity is inactivated by mutation of catalytic histidine residues in the HEPN domains (H133A/H1058A). Unknown
pC0056 - LwCas13a-msfGFP-NESLwCas13a-msfGFP-NES (Homo sapiens) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses active LwaCas13a-NES for mammalian RNA targeting lenti(AMP)
pXR001: EF1a-CasRx-2A-EGFPCasRx (Homo sapiens)EF1AGFP Hsu Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033. CasRx (NLS-RfxCas13d-NLS) with 2A-EGFP for RNA targeting lenti(AMP)

Bacterial

ID Plasmid Gene/Insert PI Publication
70168pZ003 (LshC2c2 locus)LshC2c2 full locus (Other) Zhang Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Mol Cell. 2015 Nov 5;60(3):385-97. doi: 10.1016/j.molcel.2015.10.008. Epub 2015 Oct 22.
70170pZ004 (LseC2c2 locus)LseC2c2 locus with one spacer in array (Other) Zhang Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Mol Cell. 2015 Nov 5;60(3):385-97. doi: 10.1016/j.molcel.2015.10.008. Epub 2015 Oct 22.
79150pC001 - huLshC2C2-MBP for bacterial expressionC2c2 (Other) Zhang C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573.
79151pC002 - LshC2C2 locus into pACYC184LshC2c2 locus (Other) Zhang C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573.
79152pC003 - LshC2C2 locus into pACYC184 for spacer cloningLshC2c2 locus (Other) Zhang C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573.
83482p2CT-His-MBP-Lbu_C2c2_WTLbu_C2c2_WT (Other) Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.
83483p2CT-His-MBP-Lbu_C2c2_R472A_H477ALbu_C2c2_R472A_H477A (Other) Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.
83484p2CT-His-MBP-Lbu_C2c2_R1048A_H1053ALbu_C2c2_R1048A_H1053A (Other) Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.
83485p2CT-His-MBP-Lbu_C2c2_R472A_H477A_R1048A_H1053ALbu_C2c2_R472A_H477A_R1048A_H1053A (Other) Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.
83486p2CT-His-MBP-Lse_C2c2_WTLse_C2c2_WT (Other) Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.
83487p2CT-His-MBP-Lsh_C2c2_WTLsh_C2c2_WT (Other) Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.
83494p2CT-His-MBP-Lbu_C2c2_R1079ALbu_C2c2_R1079A (Other) Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.
89898pBZCas13bCas13b (Other) Zhang Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.
89901pBZCas13b-HEPNCas13b (Other) Zhang Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.
89906pPbcas13bCas13b (Other) Zhang Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.
90097pC013 - Twinstrep-SUMO-huLwCas13aLwCas13a (Synthetic) Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
91855p2CT-His-MBP-Lbu_C2c2_E299ALbuCas13a (LbuC2c2) E299A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91856p2CT-His-MBP-Lbu_C2c2_K310ALbuCas13a (LbuC2c2) K310A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91858p2CT-His-MBP-Lbu_C2c2_N314ALbuCas13a (LbuC2c2) N314A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91859p2CT-His-MBP-Lbu_C2c2_R1072ALbuCas13a (LbuC2c2) R1072A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91860p2CT-His-MBP-Lbu_C2c2_D1078ALbuCas13a (LbuC2c2) D1078A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91861p2CT-His-MBP-Lbu_C2c2_R1079A_K1080ALbuCas13a (LbuC2c2) R0179A/K1080A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91862p2CT-His-MBP-Lbu_C2c2_K1080ALbuCas13a (LbuC2c2) K1080A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91863p2CT-His-MBP-Lbu_C2c2_K1082ALbuCas13a (LbuC2c2) K1082A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91864p2CT-His-MBP-Lbu_C2c2_K1087ALbuCas13a (LbuC2c2) K1087A (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91865p2CT-His-MBP-Lwa_Cas13a_WTLwaCas13a (LwaC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91866p2CT-His-MBP-Lne_Cas13a_WTLneCas13a (LneC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91867p2CT-His-MBP-Lba_Cas13a_WTLbaCas13a (LbaC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91868p2CT-His-MBP-Ere_Cas13a_WTEreCas13a (EreC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91869p2CT-His-MBP-Cam_Cas13a_WTCamCas13a (CamC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91870p2CT-His-MBP-Rca_Cas13a_WTRcaCas13a (RcaC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91871p2CT-His-MBP-Hhe_Cas13a_WTHheCas13a (HheC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91872p2CT-His-MBP-Ppr_Cas13a_WTPprCas13a (PprC2c2) WT (Other) Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.
91900pC009 LshCas13a locus with targeting spacerLshCas13a locus with targeting spacer (Other) Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
91901pC010 LshCas13a locus with nontargeting spacerLshCas13a locus with non-targeting spacer (Other) Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
91903pC011 LwCas13a locus with targeting spacerLwCas13a with targeting spacer (Other) Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
91904pC012 LwCas13a locus with nontargeting spacerLwCas13a with non-targeting spacer (Other) Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
91908pC018 - LshCas13a from Leptotrichia shahiiLshCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91909pC019 - LwCas13a from Leptotrichia wadeiLwCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91910pC020 - LseCas13a from Listeria seeligeriLseCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91911pC021 - LbmCas13a from Lachnospiraceae bacterium MA2020LbmCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91912pC022 - LbnCas13a from Lachnospiraceae bacterium NK4A179LbnCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91913pC023 - CaCas13a from [Clostridium] aminophilum DSM 10710CaCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91914pC024 - CgCas13a from Carnobacterium gallinarum DSM 4847CgCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91915pC025 - Cg2Cas13a from Carnobacterium gallinarum DSM 4847Cg2Cas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91916pC026 - PpCas13a from Paludibacter propionicigenes WB4PpCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91917pC027 - LweCas13a from Listeria weihenstephanensis FSL R9-0317LweCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91918pC028 - LbfCas13a from Listeriaceae bacterium FSL M6-0635LbfCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91919pC029 - Lw2Cas13a from Leptotrichia wadei F0279Lw2Cas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91920pC030 - RcsCas13a from Rhodobacter capsulatus SB 1003RcsCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91921pC031 - RcrCas13a from Rhodobacter capsulatus R121RcrCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
91922pC032 - RcdCas13a from Rhodobacter capsulatus DE442RcdCas13a and guide expression cassette (Other) Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.
100817pDuBir-Lbu-dCas13a-avitagLbu_Cas13a_R472A_H477A_R1048A_H1053A (Other) O'Connell RNA Binding and HEPN-Nuclease Activation Are Decoupled in CRISPR-Cas13a. Cell Rep. 2018 Jul 24;24(4):1025-1036. doi: 10.1016/j.celrep.2018.06.105.
108303pET28a-MH6-EsCas13dEsCas13d (Synthetic) Biotechnologies Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028.
108304pET28a-MH6-RspCas13d_RspCasWYL1RspCas13d (Synthetic), RspWYL1 (Synthetic) Biotechnologies Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028.
108305pET28a-MH6-RspCas13dRspCas13d (Synthetic) Biotechnologies Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028.


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