{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Nakahara E"],"funding":["NEI NIH HHS","National Eye Institute","Chan Zuckerberg Initiative","Roger and Dorothy Hirl Research Fund"],"pagination":["2877-2889"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11212008"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["17(10)"],"pubmed_abstract":["Destabilizing domains (DDs) are an attractive strategy allowing for positive post-transcriptional small molecule-regulatable control of a fusion protein's abundance. However, in many instances, the currently available DDs suffer from higher-than-desirable basal levels of the fusion protein. Accordingly, we redesigned the <i>E. coli</i> dihydrofolate reductase (ecDHFR) DD by introducing a library of ∼1200 random ecDHFR mutants fused to YFP into CHO cells. Following successive rounds of fluorescence-activated cell sorting, we identified six new ecDHFR DD clones with significantly enhanced proteasomal turnover in the absence of a stabilizing ligand, trimethoprim (TMP). One of these clones, designated as \"C12\", contained four unique missense mutations (W74R/T113S/E120D/Q146L) and demonstrated "],"journal":["ACS chemical biology"],"pubmed_title":["Development of a New DHFR-Based Destabilizing Domain with Enhanced Basal Turnover and Applicability in Mammalian Systems."],"pmcid":["PMC11212008"],"funding_grant_id":["R21 EY033522","R01 EY027785","EY030413","2018-191983","EY027785","P30 EY030413"],"pubmed_authors":["Collier GE","Hulleman JD","Nakahara E","Mullapudi V","Joachimiak LA"],"additional_accession":[]},"is_claimable":false,"name":"Development of a New DHFR-Based Destabilizing Domain with Enhanced Basal Turnover and Applicability in Mammalian Systems.","description":"Destabilizing domains (DDs) are an attractive strategy allowing for positive post-transcriptional small molecule-regulatable control of a fusion protein's abundance. However, in many instances, the currently available DDs suffer from higher-than-desirable basal levels of the fusion protein. Accordingly, we redesigned the <i>E. coli</i> dihydrofolate reductase (ecDHFR) DD by introducing a library of ∼1200 random ecDHFR mutants fused to YFP into CHO cells. Following successive rounds of fluorescence-activated cell sorting, we identified six new ecDHFR DD clones with significantly enhanced proteasomal turnover in the absence of a stabilizing ligand, trimethoprim (TMP). One of these clones, designated as \"C12\", contained four unique missense mutations (W74R/T113S/E120D/Q146L) and demonstrated ","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Oct","modification":"2026-05-29T15:09:06.326Z","creation":"2026-04-08T05:06:44.667Z"},"accession":"S-EPMC11212008","cross_references":{"pubmed":["36122928"],"doi":["10.1021/acschembio.2c00518"]}}