big congrats, my friend, awesome work! 🎉

congrats, Walter!!

Congrats to you and your team, Natalie!!

...as secreted by millions of other cells in the population. our prior screen data, at least, suggested that NADK2 was diff. essential in DMEM v. HPLM or RPMI regardless of FBS dialysis, though we also often use relatively lower seeding density, etc. just some speculation at least, great q! 4/4

...so, in the context of a pooled population as is the basis of pooled CRISPR screens, it is certainly possible (if not likely) that based on seeding density and passaging frequency used thru the screen duration, the NADK2-KO cells within the population may indeed be exposed to exogenous Pro...3/n

...also possibly just an artifactual consequence of technical parameters used for specific CRISPR screens in 'batch' format (though often not reported). we and others have seen that in some cases, cultured cells 'net secrete' Pro into exogenous media over time...2/n

thanks so much, Dylan!! great q! edited for clarity :)
1-for the screens carried out in RPMI, this would certainly seem to be the case (there is plenty of exogenous Pro around)
2-for those in DMEM, it may be that 'standard' 10-20% FBS provides ample Pro...but...1/n

thanks so much, Christian!! 🙏

thanks, Iza - hope that all is well!!

thank you, my friend!! 🙏

thanks, Natalie!! 🎉

thank you, Tim!!

thanks so much, Alex!! 😀

armed with his metabolism background, Kyle will now go on to pursue his postdoc exploring regen bio w/Ken Poss, while we continue our ongoing efforts to unravel mechanistic insights into conditional gene essentiality among other directions! 11/11

nonetheless, our findings indicate that selective NADK inhibitors may be leveraged to effectively slow DHFR activity, impairing cell growth while eluding resistance mechanisms related to direct DHFR inhibitors such as methotrexate...10/n

given the central role of NADPH in several processes, it comes as no surprise that NADK has garnered attention as a possible target for cancer therapy, but using NADK-KO cells, we corroborate that TN (deemed a lead compound against NADK) elicits off-target effects...9/n