Teton plko

Manufactured by Addgene

The Teton-pLKO is a plasmid designed for use in lentiviral-based gene knockdown experiments. It contains a puromycin resistance cassette and the necessary lentiviral packaging elements for production of recombinant lentiviral particles.

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Lab products found in correlation

Pcdna3.1 v5 his, by Thermo Fisher Scientific (1 mentions) Agei ecori linearized plko 1 vector, by Addgene (1 mentions)

2 protocols using teton plko

Construction and Validation of PRDX Plasmids

Cited 1 time

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The complete coding sequences of human PRDX1-6 were amplified from HeLa cell cDNA by PCR and ligated into pcDNA3.1-V5-His (Invitrogen) for transient transfection. PRDX2 or PRDX4 cDNA was also ligated into pLenti4-puro for stable transfection. The catalytically inactive PRDX2(C51S) and PRDX4(C124S) mutants were generated using QuickChange Site-directed Mutagenesis Kit (Stratagene). PRDX2 and PRDX4 shRNA oligonucleotides were ligated into Teton-pLKO (Addgene #21915). HIF-1α and HIF-2α shRNAs were cloned into pLKO.1. The oligonucleotide sequences of shRNAs are shown in Table 1. Other constructs have been described previously [41 (link), 42 (link)]. The DNA sequences of all recombinant plasmids were confirmed by nucleotide sequence analysis.

Luo W., Chen I., Chen Y., Alkam D., Wang Y, & Semenza G.L. (2016). PRDX2 and PRDX4 are negative regulators of hypoxia-inducible factors under conditions of prolonged hypoxia. Oncotarget, 7(6), 6379-6397.

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Molecular Cloning of CHD4, MBD3, MTA2, and HIF-2α

Cited 2 times

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Full-length CHD4 cDNA was generated by PCR using mouse CHD4 cDNA (Harvard plasmid, MmCD00320214) as a template and cloned into HpaI/NheI-linearized pcFUGW-3XFLAG vector. FLAG-CHD4 (K750R) was generated by site-directed PCR mutagenesis. Human MBD3 and MTA2 cDNAs were amplified from MDA-MB-321 cell cDNA by PCR and cloned into pcDNA3.1-V5 vector. cDNAs encoding human truncated HIF-2α domains were cloned to pGEX-6P-1 vector. DNA oligonucleotides of the short hairpin RNAs (shRNAs) targeting CHD4 (Table S1) were annealed and ligated into AgeI/EcoRI-linearized pLKO.1 vector (Addgene, #8435) or Teton-pLKO (Addgene, #21915). Other constructs have been described previously (14 (link),15 (link),26 (link)). The DNA sequences of all recombinant plasmids were confirmed by nucleotide sequence analysis.

Wang Y., Chen Y., Bao L., Zhang B., Wang J.E., Kumar A., Xing C., Wang Y, & Luo W. (2020). CHD4 promotes breast cancer progression as a coactivator of hypoxia-inducible factors. Cancer research, 80(18), 3880-3891.

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