A multiplexed gRNA piggyBac transposon system facilitates efficient induction of CRISPRi and CRISPRa in human pluripotent stem cells.
Hazelbaker DZ, Beccard A, Angelini G, Mazzucato P, Messana A, Lam D, Eggan K, Barrett LE
Sci Rep. 2020 Jan 20;10(1):635. doi: 10.1038/s41598-020-57500-1.
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Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 128467 | S1_AscI_V5-gateway_Bsu36I | destination vector with N-terminal V5 tag |
| 128468 | S2_AscI_HA-gateway_Bsu36I | destination vector with N-terminal HA tag |
| 128469 | S5_EcoRV_Ef1a_AscI | Ef1a promoter |
| 128470 | S6_BstBI_SV40_SwaI | SV40 promoter |
| 128471 | S7_SwaI_Zeo_PacI | Zeo Resistance gene |
| 128472 | S8_attL1-N-term-EGFP-attL2 | EGFP entry clones |
| 128473 | S9_attL1-EGFP-C-term-attL2 | EGFP entry clone |
| 128474 | S10_EcoRV_hcSYN_AscI | hcSYN promoter |
| 128475 | S11_EcoRV_CAMKIIA_AscI | CAMKII1 promoter |
| 128476 | S12_EcoRV_PGK_AscI | PGK promoter |
| 128477 | S13_EcoRV_UBC_AscI | UBC promoter |
| 137865 | pPN454 | Expression of gRNA i1 targeting TCF4 for CRISPRi; U6 promoter |
| 137866 | pPN434 | Expression of gRNA i2 targeting TCF4 for CRISPRi; U6 promoter |
| 137867 | pPN435 | Expression of gRNA i3 targeting TCF4 for CRISPRi; U6 promoter |
| 137868 | pPN446 | Expression of gRNA a1 targeting TCF4 for CRISPRa; U6 promoter |
| 137869 | pPN431 | Expression of gRNA a2 targeting TCF4 for CRISPRa; U6 promoter |
| 137870 | pPN432 | Expression of gRNA a3 targeting TCF4 for CRISPRa; U6 promoter |
| 137871 | pPN447 | Expression of gRNA targeting POGZ for CRISPRi; U6 promoter |
| 137872 | pPN455 | Expression of gRNA targeting POGZ for CRISPRa; U6 promoter |
| 137873 | pPN441 | PiggyBac vector for expression of 3xgRNA targeting TCF4 for CRISPRi; mRFP-T2A-BlasticidinR |
| 137874 | pPN440 | PiggyBac vector for expression of 3xgRNA targeting TCF4 for CRISPRa; mRFP-T2A-BlasticidinR |
| 137875 | pPN459 | PiggyBac vector for expression of 1x gRNA targeting TCF4 for CRISPRi; mRFP-T2A-BlasticidinR |
| 137876 | pPN458 | PiggyBac vector for expression of 1x gRNA targeting TCF4 for CRISPRa;mRFP-T2A-BlasticidinR |
| 137877 | pPN461 | PiggyBac vector for expression of 1x gRNA targeting POGZ for CRISPRi; mRFP-T2A-BlasticidinR |
| 137878 | pPN460 | PiggyBac vector for expression of 1x gRNA targeting POGZ for CRISPRa;mRFP-T2A-BlasticidinR |
| 137879 | pT077 | Donor vector for integration into the human AAVS1 safe harbor locus of Doxicycline-inducible KRAB-dCas9-IRES-EGFP |
| 137880 | pT076 | Donor vector for integration at the human AAVS1 safe harbor locus of Doxicycline-inducible dCas9-VPR-T2A-EGFP |
| 137881 | pGEP163 | PiggyBac donor vector for expression of mRFP-T2A-BlasticidinR |
| 137882 | pGEP179_pX330K | Entry vector for multi-gRNA cloning |