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intramural research program. She inquired about the engineering of an immortalized cell line with a key protein knocked out.This
cell line will be used in the Mindell lab to to study CLC7 and lysosomal pH regulation. She discussed the design of the cells,
timeline for production and cost with numerous vendors. She narrowed down the options based on what was discussed and then
obtained quotes from Synthego, HD Biosciences and SystemBio.
Our goal in the Mindell lab is to better understand lysosomal pH regulation. The signaling lipid PI(3,5)P2 modulates lysosomal dynamics, including by regulating lysosomal ion channels, raising the possibility that it could contribute to lysosomal pH regulation. We have demonstrated that depleting PI(3,5)P2 by inhibiting the kinase PIKfyve causes lysosomal hyperacidification, primarily via an effect on ClC-7. We have also found that PI(3,5)P2 directly inhibits ClC-7 transport and that this inhibition is eliminated in a disease-causing gain-of-function ClC-7 mutation. Together, these observations suggest an intimate role for ClC-7 in lysosomal pH regulation.These two immortalized cell lines will allow us to evaluate the effect of this lysosomal regulating lipid (PI35P2) on the chloride/proton antiporter CLC-7 by deleting an enzyme responsible for regulating it.
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