An expanded toolkit of drug resistance cassettes for Candida glabrata , Candida auris , and Candida albicans leads to new insights into the ergosterol pathway.

Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 101674533 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2379-5042 (Electronic) Linking ISSN: 23795042 NLM ISO Abbreviation: mSphere Subsets: MEDLINE

Original Publication: Washington, DC : American Society for Microbiology, [2015]-

Candida albicans*/genetics ; Candida glabrata*; Antifungal Agents/pharmacology ; Antifungal Agents/metabolism ; Candida auris ; Ergosterol ; Candida/genetics ; Drug Resistance, Fungal/genetics

Importance: The increasing problem of drug resistance and emerging pathogens is an urgent global health problem that necessitates the development and expansion of tools for studying fungal drug resistance and pathogenesis. Prior studies in Candida glabrata , Candida auris , and Candida albicans have been mainly limited to the use of NatMX/SAT1 and HphMX/CaHyg for genetic manipulation in prototrophic strains and clinical isolates. In this study, we demonstrated that NatMX/SAT1, HphMX, KanMX, and/or BleMX drug resistance cassettes when coupled with a CRISPR-ribonucleoprotein (RNP)-based system can be efficiently utilized for deleting or modifying genes in the ergosterol pathway of C. glabrata , C. auris , and C. albicans . Moreover, the utility of these tools has provided new insights into ERG genes and their relationship to azole resistance in Candida . Overall, we have expanded the toolkit for Candida pathogens to increase the versatility of genetically modifying complex pathways involved in drug resistance and pathogenesis.
Competing Interests: The authors declare no conflict of interest.

Biochem Biophys Res Commun. 1989 Nov 15;164(3):1170-5. (PMID: 2556119)
Yeast. 1998 Jul;14(10):943-51. (PMID: 9717240)
J Biol Chem. 2022 Oct;298(10):102485. (PMID: 36108742)
Open Forum Infect Dis. 2018 Jul 31;5(8):ofy187. (PMID: 30151412)
Fungal Genet Biol. 2010 Feb;47(2):81-93. (PMID: 19595784)
mSphere. 2016 Jun 15;1(3):. (PMID: 27340698)
Nucleic Acids Res. 1996 Jul 1;24(13):2519-24. (PMID: 8692690)
mBio. 2019 Jan 29;10(1):. (PMID: 30696744)
Antimicrob Agents Chemother. 2019 Mar 27;63(4):. (PMID: 30718246)
Antimicrob Agents Chemother. 2022 May 17;66(5):e0225021. (PMID: 35471041)
J Biol Chem. 1997 Apr 11;272(15):9986-8. (PMID: 9092539)
mSphere. 2017 Jun 21;2(3):. (PMID: 28657070)
Yeast. 2012 Mar;29(3-4):119-36. (PMID: 22473760)
FEMS Yeast Res. 2017 Sep 1;17(6):. (PMID: 28910986)
Microb Pathog. 2021 Mar;152:104616. (PMID: 33212195)
Sci Rep. 2016 Oct 21;6:35766. (PMID: 27767081)
mSphere. 2022 Aug 31;7(4):e0034722. (PMID: 35968963)
Antimicrob Agents Chemother. 2002 Jun;46(6):1704-13. (PMID: 12019079)
Genetics. 1999 Mar;151(3):979-87. (PMID: 10049916)
Med Mycol. 2005 Jun;43(4):285-318. (PMID: 16110776)
Curr Genet. 2005 Aug;48(2):117-25. (PMID: 16078083)
Yeast. 2017 May;34(5):223-235. (PMID: 27943405)
Clin Microbiol Rev. 2007 Jan;20(1):133-63. (PMID: 17223626)
Sci Adv. 2015;1(3):e1500248. (PMID: 25977940)
Microbiol Spectr. 2022 Jun 29;10(3):e0263021. (PMID: 35612314)
Genetics. 1993 Jul;134(3):717-28. (PMID: 8349105)
Antimicrob Agents Chemother. 1995 Dec;39(12):2708-17. (PMID: 8593007)
PLoS Genet. 2018 Apr 27;14(4):e1007319. (PMID: 29702647)
PLoS Pathog. 2014 Jun 19;10(6):e1004211. (PMID: 24945925)
Trends Mol Med. 2002 Feb;8(2):76-81. (PMID: 11815273)
Microbiol Immunol. 2009 Jan;53(1):41-4. (PMID: 19161556)
Antimicrob Agents Chemother. 2010 Nov;54(11):4527-33. (PMID: 20733039)
FEBS Lett. 1996 Aug 26;392(2):161-5. (PMID: 8772195)
Front Microbiol. 2013 May 29;4:127. (PMID: 23755044)
Curr Biol. 2021 Dec 6;31(23):R1520-R1523. (PMID: 34875240)
J Biol Chem. 2005 Aug 5;280(31):28761-5. (PMID: 15964832)
mBio. 2018 Jul 24;9(4):. (PMID: 30042199)
Mol Cell Biol. 2010 Jul;30(13):3216-32. (PMID: 20439497)
Open Forum Infect Dis. 2022 Jan 10;9(1):ofab593. (PMID: 35036461)
mSphere. 2018 Apr 25;3(2):. (PMID: 29695624)
Antimicrob Agents Chemother. 2003 Aug;47(8):2404-12. (PMID: 12878497)
Microb Cell. 2022 Oct 13;9(11):174-189. (PMID: 36448018)
Biochem J. 1990 Mar 1;266(2):475-80. (PMID: 2180400)
Curr Biol. 2021 Dec 6;31(23):5314-5326.e10. (PMID: 34699784)
mBio. 2019 Jun 11;10(3):. (PMID: 31186322)
Mycoses. 2006 Nov;49(6):445-51. (PMID: 17022759)
mSphere. 2023 Oct 24;8(5):e0028323. (PMID: 37594261)
Clin Microbiol Infect. 2022 Jun;28(6):838-843. (PMID: 34915074)
Gene. 1996 Feb 22;169(1):105-9. (PMID: 8635732)
J Biol Chem. 2012 Jan 20;287(4):2652-65. (PMID: 22147691)
Antimicrob Agents Chemother. 1999 Jul;43(7):1725-8. (PMID: 10390230)
Yeast. 1999 Oct;15(14):1541-53. (PMID: 10514571)
J Biol Chem. 2010 Jan 1;285(1):595-607. (PMID: 19897479)
Gene. 2004 Oct 27;341:119-27. (PMID: 15474295)
Front Microbiol. 2013 Jan 10;3:439. (PMID: 23335918)
Eur J Clin Microbiol Infect Dis. 2014 May;33(5):673-88. (PMID: 24249283)
Genes (Basel). 2020 Jul 15;11(7):. (PMID: 32679672)
J Fungi (Basel). 2017 Oct 18;3(4):. (PMID: 29371573)
Yeast. 2020 Jan;37(1):27-44. (PMID: 31800968)

R01 AI136995 United States AI NIAID NIH HHS; T32 AI148103 United States AI NIAID NIH HHS

Keywords: CRISPR-RNP; Candida albicans; Candida auris; Candida glabrata; ERG5 and ERG3; KanMX and BleMX; antifungal drug resistance; azole antifungal drugs; ergosterol pathway; toxic sterols

0 (Antifungal Agents)
Z30RAY509F (Ergosterol)

Date Created: 20231106 Date Completed: 20231221 Latest Revision: 20231222

20231222

PMC10732037

10.1128/msphere.00311-23

37929964