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Proteome characterization of XPC-deficient melanocytes generated by CRISPR-Cas9 technology reveals alteration in the expression of several hundred of proteins

ABSTRACT: Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder owing to the defects in one of the factors implicated in nucleotide excision repair (NER) system. The effect of NER deficiency on melanocyte biology has not received much attention up to now from a mechanistic standpoint, due to the lack of an appropriate model. Therefore, we decided to generate XPC-deficient primary melanocytes by CRISPR-Cas9 technology, herein after called ©-XPC melanocytes. To this end, we designed two gRNA for exon 9 close to the most common XPC mutation (i.e. c.1643_1644delTG). These gRNA were cloned into lentiviral vectors containing the Cas9 nuclease and puromycin selection cassettes. Following transduction with lentivirus, melanocytes sustainably expressing Cas-9 and scramble gRNA (©-Ctrl) or gRNA targeting XPC gene (©-XPC) were subjected to PCR analysis, Sanger sequencing and western blotting. Following validation of our model, ©-Ctrl and ©-XPC melanocytes were subjected to a quantitative label-free differential proteomic analysis.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Dupuy Jean-William

LAB HEAD: Hamid Reza Rezvani

PROVIDER: PXD042713 | Pride | 2024-12-17

REPOSITORIES: Pride

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	Link_between_files.xlsx	Xlsx
	RP_UP000005640_Homo_sapiens_190516.fasta	Fasta
	checksum.txt	Txt
	mc190514_01.raw	Raw
	mc190514_01.xlsx	Xlsx

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Proteome characterization of XPC-deficient melanocytes generated by CRISPR-Cas9 technology reveals alteration in the expression of several hundred proteins.

Cario Muriel M Scalia Julie J Mahfouf Walid W Muzotte Elodie E Michaud Vincent V Plaisant Claudio C Dupuy Jean-William JW Douki Thierry T Morice-Picard Fanny F Rambert Jérôme J Perrier Eric E Trompezinski Sandra S Rezvani Hamid-Reza HR

Journal of dermatological science 20240323 2

PMID: 38556435

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