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Human gatad2a orf

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The Human GATAD2A ORF is a DNA construct containing the full-length coding sequence for the GATAD2A (GATA Zinc Finger Domain Containing 2A) gene from Homo sapiens. GATAD2A is a protein that functions as a transcriptional regulator, involved in chromatin remodeling and gene expression.

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Lab products found in correlation

Plvx puro vector, by Takara Bio (2 mentions) Phr sffv dcas9 bfp krab, by Addgene (2 mentions)

2 protocols using human gatad2a orf

1

Overexpression of GATAD2A Constructs

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Human GATAD2A ORF was purchased from GenScript (Clone ID: OHu17401D, RefSeq: NM_017660.3) and was PCR amplified and cloned between EcoRI and BamHI restriction sites of a pLVX-Puro vector (Clontech) using Gibson assembly with primers provided in Supplementary Table 3. To generate SFFV promoter driven GATAD2A 335–486 construct, CMV promoter of pLVX-puro was replaced with SFFV promoter (amplified from pHR-SFFV-dCas9-BFP-KRAB, Addgene #46911). Briefly, pLVX-puro GATAD2A 335–486 was digested with ClaI and EcoRI, and the large vector fragment was gel purified for further Gibson assembly with PCR amplified SFFV promoter. The sequence integrity of all the cloned products were verified by Sanger sequencing. All the cloned GATAD2A constructs contain a SV-40 NLS (PKKKRKV) and a FLAG-tag (DYKDDDDK) at the N-terminus. Wild type HUDEP-2 cells were transduced with lentivirus carrying GATAD2A constructs for 24 hours, followed by selection with puromycin (1 μg/ml).
Sher F., Hossain M., Seruggia D., Schoonenberg V.A., Yao Q., Cifani P., Dassama L.M., Cole M.A., Ren C., Vinjamur D.S., Macias-Trevino C., Luk K., McGuckin C., Schupp P.G., Canver M.C., Kurita R., Nakamura Y., Fujiwara Y., Wolfe S.A., Pinello L., Maeda T., Kentsis A., Orkin S.H, & Bauer D.E. (2019). Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis. Nature genetics, 51(7), 1149-1159.
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2

Overexpression of GATAD2A Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human GATAD2A ORF was purchased from GenScript (Clone ID: OHu17401D, RefSeq: NM_017660.3) and was PCR amplified and cloned between EcoRI and BamHI restriction sites of a pLVX-Puro vector (Clontech) using Gibson assembly with primers provided in Supplementary Table 3. To generate SFFV promoter driven GATAD2A 335–486 construct, CMV promoter of pLVX-puro was replaced with SFFV promoter (amplified from pHR-SFFV-dCas9-BFP-KRAB, Addgene #46911). Briefly, pLVX-puro GATAD2A 335–486 was digested with ClaI and EcoRI, and the large vector fragment was gel purified for further Gibson assembly with PCR amplified SFFV promoter. The sequence integrity of all the cloned products were verified by Sanger sequencing. All the cloned GATAD2A constructs contain a SV-40 NLS (PKKKRKV) and a FLAG-tag (DYKDDDDK) at the N-terminus. Wild type HUDEP-2 cells were transduced with lentivirus carrying GATAD2A constructs for 24 hours, followed by selection with puromycin (1 μg/ml).
Sher F., Hossain M., Seruggia D., Schoonenberg V.A., Yao Q., Cifani P., Dassama L.M., Cole M.A., Ren C., Vinjamur D.S., Macias-Trevino C., Luk K., McGuckin C., Schupp P.G., Canver M.C., Kurita R., Nakamura Y., Fujiwara Y., Wolfe S.A., Pinello L., Maeda T., Kentsis A., Orkin S.H, & Bauer D.E. (2019). Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis. Nature genetics, 51(7), 1149-1159.
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