High-Throughput Screens of PAM-Flexible Cas9 Variants for Gene Knockout and Transcriptional Modulation.
Legut M, Daniloski Z, Xue X, McKenzie D, Guo X, Wessels HH, Sanjana NE
Cell Rep. 2020 Mar 3;30(9):2859-2868.e5. doi: 10.1016/j.celrep.2020.02.010.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 139086 | pCC_01 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-Cas9-NLS-2A-Puro-WPRE | Expresses human codon-optimized SpCas9 nuclease in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a unique barcode downstream of sgRNA cassette. |
| 139087 | pCC_02 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-Cas9NG-NLS-2A-Puro-WPRE | Expresses human codon-optimized Cas9-NG nuclease in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a unique barcode downstream of sgRNA cassette. |
| 139088 | pCC_03 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-xCas9-NLS-2A-Puro-WPRE | Expresses human codon-optimized xCas9 3.7 nuclease in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a unique barcode downstream of sgRNA cassette. |
| 139089 | pCC_04 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-xCas9NG-NLS-2A-Puro-WPRE | Expresses human codon-optimized xCas9-NG nuclease in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a unique barcode downstream of sgRNA cassette. |
| 139090 | pCC_05 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-dCas9-NLS-VPR-2A-Puro-WPRE | Expresses human codon-optimized inactive SpCas9 fused to a transcriptional activator VPR in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |
| 139091 | pCC_06 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-dCas9NG-NLS-VPR-2A-Puro-WPRE | Expresses human codon-optimized inactive SpCas9-NG fused to a transcriptional activator VPR in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |
| 139092 | pCC_07 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-dxCas9-NLS-VPR-2A-Puro-WPRE | Expresses human codon-optimized inactive xCas9 3.7 fused to a transcriptional activator VPR in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |
| 139093 | pCC_08 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-dxCas9NG-NLS-VPR-2A-Puro-WPRE | Expresses human codon-optimized inactive xCas9-NG fused to a transcriptional activator VPR in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |
| 139094 | pCC_09 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-KRAB-dCas9-NLS-2A-Puro-WPRE | Expresses human codon-optimized inactive SpCas9 fused to a transcriptional repressor KRAB in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |
| 139095 | pCC_10 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-KRAB-dCas9NG-NLS-2A-Puro-WPRE | Expresses human codon-optimized inactive SpCas9-NG fused to a transcriptional repressor KRAB in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |
| 139096 | pCC_11 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-KRAB-dxCas9-NLS-2A-Puro-WPRE | Expresses human codon-optimized inactive xCas9 3.7 fused to a transcriptional repressor KRAB in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |
| 139097 | pCC_12 - hU6-BsmBI-sgRNA(E+F)-barcode-EFS-KRAB-dxCas9NG-NLS-2A-Puro-WPRE | Expresses human codon-optimized inactive xCas9-NG fused to a transcriptional repressor KRAB in mammalian cells. For cloning of sgRNAs using BsmBI. Contains a barcode downstream of sgRNA cassette. |