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Hi, this is my first tweet to test the system. I plan to share some of my lab experiments/struggles with you. At the moment, I am trying to knockout the PKD1 gene in its entirety in RCTE cells. I think I am getting close and will tweet once I get proof that I have achieved it.

Think I managed to delete the entire PKD1 gene in human RCTE cells. Looks like two alleles in this cell line. Will be useful as a negative control in IF.

The Cas12a system works very well (thank you IDT!). There is a video protocol showing the approach at youtube.com/watch?v=WoJZED… . Next up some IF!

Hi, the Western blots are to show that in the D_A cell line (with the deletion of the entire PKD1 gene) we don't have the high mol wt 7E12 (anti-polycystin-1) bands, seen WT cells. The other blots are to show that the Westerns were equally loaded between WT and D_A. So 7E12 works

Therefore, we have a null human cell line and an antibody that is mostly specific. I hope that explains things a bit better. We need to check by immunofluorescence next.

Just finished editing some new technical videos on YouTube. Interesting if you want to know how to make monoclonal antibodies or edit a large piece of DNA. youtube.com/@christopherwa…

Here's something pretty! Ulex europaeus agglutinin (UEA, fucose specific) -red on a pig kidney section. Seems specific for the thick ascending limb of the LOH. In humans UEA detects vascular endothelium-- but not in the pig. A bit of new information!

Hi, new paper describing the interaction between CU062 and polycystin-1. Three new mAbs, one to PC2 and two to CU062, new knockout cell lines for PKD1 and PKD2 in RCTE cells. Suggests a solution to the finding of mitochondrial defects in ADPKD. mdpi.com/2073-4409/12/1…