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Moreover, homozygous deletion of non-essential genes has often been problematic due to the frequent aneuploidy in the mutant strains. Rapid, conditional depletion of essential genes by the anchor-away strategy has been successfully employed in\n              <jats:italic>Saccharomyces cerevisiae<\/jats:italic>\n              and other model organisms. Here, rapamycin mediates the dimerization of human FK506-binding protein (FKBP12) and FKBP12-rapamycin-binding (FRB) domain-containing target protein, resulting in relocalization to altered sub-cellular locations. In this work, we used the ribosomal protein Rpl13 as the anchor and took two nuclear proteins as targets to construct a set of mutants in a proof-of-principle approach. We first constructed a rapamycin-resistant\n              <jats:italic>C. albicans<\/jats:italic>\n              strain by introducing a dominant mutation in the\n              <jats:italic>CaTOR1<\/jats:italic>\n              gene and a homozygous deletion of\n              <jats:italic>RBP1<\/jats:italic>\n              , the ortholog of\n              <jats:italic>FKBP12<\/jats:italic>\n              , a primary target of rapamycin. The\n              <jats:italic>FKBP12<\/jats:italic>\n              and the\n              <jats:italic>FRB<\/jats:italic>\n              coding sequences were then CUG codon-adapted for\n              <jats:italic>C. albicans<\/jats:italic>\n              by site-directed mutagenesis. Anchor-away strains expressing the essential\n              <jats:italic>TBP1<\/jats:italic>\n              gene or the non-essential\n              <jats:italic>SPT8<\/jats:italic>\n              gene as\n              <jats:italic>FRB<\/jats:italic>\n              fusions were constructed. We found that rapamycin caused rapid cessation of growth of the TBP-AA strain within 15 minutes and the SPT8-AA strain phenocopied the constitutive filamentous phenotype of the\n              <jats:italic>spt8<\/jats:italic>\n              \u0394\/\n              <jats:italic>spt8<\/jats:italic>\n              \u0394 mutant. Thus, the anchor-away toolbox for\n              <jats:italic>C. albicans<\/jats:italic>\n              developed here can be employed for genome-wide analysis to identify gene function in a rapid and reliable manner, further accelerating anti-fungal drug development in\n              <jats:italic>C. albicans<\/jats:italic>\n              .\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              Molecular genetic studies thus far have identified ~27% open-reading frames as being essential for the vegetative growth of\n              <jats:italic>Candida albicans<\/jats:italic>\n              in rich medium out of a total 6,198 haploid set of open reading frames. However, a major limitation has been to construct rapid conditional alleles of essential\n              <jats:italic>C. albicans<\/jats:italic>\n              genes with near quantitative depletion of encoded proteins. Here, we have developed a toolbox for rapid and conditional depletion of genes that would aid studies of gene function of both essential and non-essential genes.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/msphere.00703-23","type":"journal-article","created":{"date-parts":[[2024,1,22]],"date-time":"2024-01-22T14:00:32Z","timestamp":1705932032000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["A CUG codon-adapted anchor-away toolkit for functional analysis of genes in\n            <i>Candida albicans<\/i>"],"prefix":"10.1128","volume":"9","author":[{"given":"Basharat Bashir","family":"Teli","sequence":"first","affiliation":[{"name":"Laboratory of Eukaryotic Gene Regulation, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 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