MCF10A cells (product no: CRL 10317), a non tumorigenic mammary epithelial cell line, and the derived isogenic line with CDH1 knock out (MCF10A CDH1-/-) using CompoZr ZFN technology (product no: CLLS1042) were purchased from Sigma. The MCF10A isogenic lines were cultured in DMEM/F12: (1:1) (Invitrogen) with 5% horse serum (Invitrogen), 10 μg/ml Actrapid Penfil neutral insulin (Novo Nordisk Pharmaceuticals Ltd), 20 ng/ml human epidermal growth factor (Peprotech), 100 ng/ml cholera toxin, and 500 ng/ml hydrocortisone (Sigma) [21 (link)]. Cells were grown at 37°C with 5% CO2, seeded into T75 flasks at densities of 3.0 × 105 and 4.5 × 105, respectively and passaged at 90% confluency (~3 days) for a maximum of ten passages (http://brugge.med.harvard.edu/protocols ).
Compozr zfn technology
Manufactured by Merck Group
Sourced in United States
CompoZr ZFN technology is a gene editing tool developed by Merck Group. It utilizes zinc finger nucleases (ZFNs) to enable precise and targeted modifications of the genome. The core function of this technology is to facilitate direct genome editing by introducing specific double-strand breaks in DNA sequences, allowing for the insertion, deletion, or alteration of genetic material.
Automatically generated - may contain errors
Lab products found in correlation
Horse serum, by Thermo Fisher Scientific (1 mentions)
Human epidermal growth factor (hegf), by Thermo Fisher Scientific (1 mentions)
Mcf10a cdh1, by Merck Group (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Cholera toxin, by Merck Group (1 mentions)
2 protocols using compozr zfn technology
1
Isogenic Cell Line Generation and Culture
2
Generation of Cre Knock-in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The donor plasmid pPPY/NLS-Cre contained a nuclear translocation signal (NLS)-cre and three SV40 polyadenylation signals (a kind gift from Dr. Gu [19] ). The targeting strategy is shown in Fig. 1. To target the mouse Ppy gene (gene ID: 19064), zinc finger nucleases (ZFN) technology has been used. ZFN plasmid design and validation was performed by CompoZr ZFN technology (Sigma-Aldrich, St. Louis, MO, USA). The ZFN were designed to cut the [TAGGTA] sequence located at the 3' end of exon 2 (Fig. 1A). The Ppy-targeting ZFN and the knock-in (KI) donor plasmid were injected into embryos, which were subsequently implanted into pseudopregnant female mice. A total of 81 pups were born.
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