RNA editing with CRISPR-Cas13.
Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung J, Zhang F
Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 103849 | pC0039-CMV-dPspCas13b-GS-ADAR2DD(E488Q) | dPspCas13b-ADAR2DD(E488Q) fusion that can be used to selectively edit adenosine to inosine in RNA molecules when used in conjuction with a guide RNA. |
| 103851 | pC0040-LwaCas13a crRNA backbone | Backbone for cloning guides that are compatible with LwaCas13a. Contains a 5' direct repeat. Clone using BbsI (BpiI). F overhang AAAC. R overhang AAAA |
| 103852 | pC0041-RanCas13b crRNA backbone | For cloning of guide RNAs compatible with RanCas13b. Contains a 3' direct repeat. Clone using BbsI (BpiI). F overhang cacc. R overhang caac. |
| 103853 | pC0042-PguCas13b crRNA backbone | For cloning of guide RNAs compatible with PguCas13b. Contains a 3' direct repeat. Clone using BbsI (BpiI). F overhang cacc. R overhang caac |
| 103854 | pC0043-PspCas13b crRNA backbone | For cloning of guide RNAs compatible with PspCas13b. Contains a 3' direct repeat. Clone using BbsI (BpiI). F overhang cacc. R overhang caac |
| 103855 | pC0044-EF1a-RanCas13b-NES-mapk | Expresses RanCas13b in mammalian cells for knockdown of target RNAs in combination with compatible guide RNAs. Localized to the cytoplasm |
| 103861 | pC0045-EF1a-PguCas13b-NES-HIV | Expresses PguCas13b in mammalian cells for knockdown of target RNAs in combination with compatible guide RNAs. Localized to the cytoplasm |
| 103862 | pC0046-EF1a-PspCas13b-NES-HIV | Expresses PspCas13b in mammalian cells for knockdown of target RNAs in combination with compatible guide RNAs. Localized to the cytoplasm |
| 103863 | pC0047-CMV-dPspCas13b-ADAR1DD(E1008Q) | dPspCas13b-ADAR1DD(E1008Q) fusion that can be used to selectively edit adenosine to inosine in RNA molecules when used in conjuction with a guide RNA |
| 103864 | pC0048-CMV-dPspCas13b-longlinker-ADAR2DD(E488Q) | dPspCas13b-ADAR2DD(E488Q) fusion that can be used to selectively edit adenosine to inosine in RNA molecules when used in conjuction with a guide RNA. Longer linker than pC0039. |
| 103865 | pC0049-EF1a-dPSPCas13b-NES-HIV, H133A/H1058A | Catalytically inactive PSPCas13b. RNase activity is inactivated by mutation of catalytic histidine residues in the HEPN domains (H133A/H1058A). |
| 103866 | pC0050-CMV-dPspCas13b-longlinker-ADAR2DD(wt) | dPspCas13b-ADAR2DD(wt) fusion that can be used to selectively edit adenosine to inosine in RNA molecules when used in conjuction with a guide RNA. Longer linker than pC0039. |
| 103867 | pC0051-W85X REPAIR targeting guide cloned into pC0043 | Guide RNA expression plasmid for dPspCas13b that can be used to direct ADAR deaminase domain activity to the W85X mutation in the luciferase reporter plasmid pC0038 |
| 103868 | pC0052-REPAIR non-targeting guide clone into pC0043 | Non-targeting guide RNA for use with PspCas13b. Useful as a control for RNA editing and RNA knockdown experiments. |
| 103869 | pC0053-CMV-dPspCas13b-GS-ADAR2DD(E488Q)-delta-984-1090 | dPspCas13b-ADAR2DD(E488Q) fusion that can be used to selectively edit adenosine to inosine in RNA molecules when used in conjuction with a guide RNA. Cas13b is truncated to allow for expression by AAV |
| 103870 | pC0054-CMV-dPspCas13b-longlinker-ADAR2DD(E488Q/T375G) | High specificity version of dPspCas13b-ADAR2DD(E488Q) fusions. T375G mutation in the ADAR deaminase domain confers increased specificity with slightly reduced activity. |
| 103871 | pC0055-CMV-dPspCas13b-GS-ADAR2DD(E488Q/T375G)-delta-984-1090 | dPspCas13b-ADAR2DD(E488Q/T375G) fusion that can be used to selectively edit adenosine to inosine in RNA when used in conjuction with a guide RNA. Cas13b is truncated to allow for expression by AAV |
| 181934 | pC0037 | Dual secreted luciferase reporter (EF1alpha-Gaussia luciferase, CMV-Cypridina luciferase) |