293T cell line has its advantages like fast growth speed and high transfection efficienc ,so we ues it much in our experiment ,but I don't know the specificity of U2OS cell line.As many cell line ,especially GFP expression related cell line use U2OS cell,I want to know the reason.Waiting for your answers
U2OS cell lines have many similar characteristics to 293T cells, fast growth and high transfection efficiencies, although maybe not the same extent. In addition to being of a different cell type (bone cancer...osteocarcinma) they do not have any adenoviral infections and therefore are a better line to perform certain experiments, and specifically no large T antigen. That being said, remember that the U2OS is a cancer cell line and will undoubtedly have mutations in cell pathways that are normally targeted by large T antigen.
That being said, a U2os will be a good line to use for many things when you want to use something other than 293
U2OS cell lines have many similar characteristics to 293T cells, fast growth and high transfection efficiencies, although maybe not the same extent. In addition to being of a different cell type (bone cancer...osteocarcinma) they do not have any adenoviral infections and therefore are a better line to perform certain experiments, and specifically no large T antigen. That being said, remember that the U2OS is a cancer cell line and will undoubtedly have mutations in cell pathways that are normally targeted by large T antigen.
That being said, a U2os will be a good line to use for many things when you want to use something other than 293
Thank you for your answer,it really solves my confusion in part. Recently ,I read paper about HR .I find many researchers use DR-GFP U2OS cell line to detect HR efficiency.I want to reconstruct a DR-GFP cell line in 293T ,so I need to know the differences between these two cell lines before I design the primer
Yes, you can use 293cells for that....I think that it's been done already, but for what you want to do, it can work.
Quote from Sotiriou SK et al., 2016 (DOI: 10.1016/j.molcel.2016.10.038),
"In an effort to identify genes that function in BIR in human cells,
we performed an siRNA screen targeting about 70 genes that
had previously been linked to DNA DSB repair and HR (Table
S1). The requirement of these genes in BIR was examined using
U2OS cells that overexpress cyclin E in an inducible manner
(Bartkova et al., 2005). In these well-characterized cells, cyclin
E overexpression leads to DRS, and the damaged replication
forks are repaired to a significant degree by BIR. Thus, when
BIR is inhibited—for example, by depleting PolD3—progression
through the cell cycle is delayed (Costantino et al., 2014)."
Hence in studying HR and DDR, U2OS cells serve as a well-characterized model. And in this case, absence of other oncogenes such as large T antigen seems important.
Moreover, Concerning telomere maintenance, U2OS cells use ALT (alternative lengthening of telomeres) while 293 cells use telomerase. see table 2 of Patrick Ryan Potts et al., 2007 (http://www.nature.com/nsmb/journal/v14/n7/pdf/nsmb1259.pdf)
Yes after the second injection
Ref:http://www.nature.com/labinvest/journal/v91/n8/full/labinvest201172a.html?foxtrotcallback=true
I have the same question with Mingjun Jiang. Why are NHEJ & HR I-SceI/ GFP cassettes constructed soly in U2OS cell line, or perhaps in 293 cell in few studies? And why only the clone with single copy integration of NHEJ or HR GFP integration was used in the studies? Can them be constructed in other (cancer) cell lines, such as Hela, A549, HepG2 and so on?
Yes, you can construct I-Sce1/GFP cassettes into HeLa cell lines, that has been done before
I have a question about U2OS drGFP strain. I would like to know the integration site of drGFP in U2OS cell line.
Please let me know where I can find the information of integration locus.
U2OS cells are much more adherent than HEK 293 and 293T cells. As such these are better suited for immunofluorescence studies when compared to HEK which loosely attach to coated cover slips. However, U2OS cells are cancerous cells with abnormalities.
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