With my experience, I noticed that high glucose concentration (45mM to be specific) does not significantly up-regulate SGLT-2 expression using western blotting technique. Can someone please suggest a way to stimulate the up-regulation of SGLT-2 (SLC5A2) expression in normal human kidney cells? If there is any protocol for this, I would appreciate any attachment of such.
I have this paper already Beatrice. Thank you anyway. I would prefer a suggestion or an attachment that would give a protocol of how to go about it not just a publication that couldn't touch my specific line of question.
Thank you Luciano, could you please provide me the protocol?
Perhaps you find something useful in these papers:
Research Engineering The Mammalian Genome With CRISPR
Article CRISPR-ERA: a comprehensive design tool for CRISPR-mediated ...
Article The new state of the art: CRISPR for gene activation and repression
One option which may work would be designing a series of sgRNA sequences which would target an dCas9-activator transgene near the promoter of your target gene. An example of this can be seen at Maeder et al., 2013, they outline their specific protocol for the up regulation of the VEGFA gene.
The authors also HEK293 cells, their protocol is detailed in the methods section of their paper (ref above).
You can induce diabetes with aloxan or streptozocin in rats to increase SGLT2 in kidney.
Thank you Robinson but my question was directed specifically to cell lines and not animal studies.
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