Título:	Identification and validation of tissue-specific housekeeping markers for the Amazon River prawn Macrobrachium amazonicum (Heller, 1862).
Autoria:	LIMA, G. M. de RODRIGUES, M. A. L. PAIXÃO, R. V. LUTZ, Í. AVIZ, M. A. B. SOUSA, J. do S. A. da L. MACIEL, B. R. QUEIROZ, L. D. MACIEL, C. M. T. SAMPAIO, I. VARELA, E. S. MACIEL, C. R.
Afiliação:	GABRIEL MONTEIRO DE LIMA, UNIVERSIDADE FEDERAL DO PARÁ; MÔNICA ANDRESSA LEITE RODRIGUES, UNIVERSIDADE FEDERAL DO PARÁ; RÔMULO VEIGA PAIXÃO; ÍTALO LUTZ, UNIVERSIDADE FEDERAL DO PARÁ; MANOEL ALESSANDRO BORGES AVIZ, INSTITUTO FEDERAL DE EDUCAÇÃO, CIÊNCIA E TECNOLOGIA DO PARÁ; JANIELI DO SOCORRO AMORIM DA LUZ SOUSA, UNIVERSIDADE FEDERAL DO PARÁ; BRUNA RAMALHO MACIEL, UNIVERSIDADE FEDERAL DO PARÁ; LUCIANO DOMINGUES QUEIROZ, INSTITUTO FEDERAL DE EDUCAÇÃO, CIÊNCIA E TECNOLOGIA DO PARÁ; CARLOS MURILO TENÓRIO MACIEL, UNIVERSIDADE FEDERAL DO PARÁ; IRACILDA SAMPAIO, UNIVERSIDADE FEDERAL DO PARÁ; EDUARDO SOUSA VARELA, CNPASA; CRISTIANA RAMALHO MACIEL, UNIVERSIDADE FEDERAL DO PARÁ.
Ano de publicação:	2025
Referência:	Genes, v. 17, n. 1, article number 26, 2025.
Conteúdo:	Background/Objectives: The selection and validation of species-specific housekeeping genes (HKGs) have become increasingly common in functional genomics, with application of quantitative Polymerase Chain Reaction (qPCR) or cDNA-based qPCR (RT-qPCR). Despite the Macrobrachium amazonicum having RNA-seq studies available, there are still no data on the most stable and consistent HKGs for use in relative gene expression analyses. Therefore, the present study aimed to identify and validate seven HKGs in M. amazonicum: Eukaryotic Translation Initiation Factor (EIF), 18S ribosomal RNA (18S), Ribosomal Protein L18 (RPL18), β-actin, α-tubulin (α-tub), Elongation Factor 1-α (EF-1α), and Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH). Methods: The HKGs were identified in the M. amazonicum transcriptome, characterized for identity confirmation, and compared against public databases. Subsequently, RT-qPCR assays were prepared using muscle, hepatopancreas, gills, testis, androgenic gland, and ovary to assess the stability of the HKG markers, employing the comparative ∆Ct, BestKeeper, NormFinder, and GeNorm methods. Results: All candidate HKGs identified showed high similarity with other decapods. Reactions performed with these markers demonstrated high specificity, PCR efficiency, and elevated coefficients of determination. The comprehensive ranking, indicated that no single HKG was stable across all tissues, with HKGs showing the best stability being tissue-specific. The most stable HKGs were RPL18 and 18S. GAPDH, historically used as an HKG, showed the poorest performance in stability ranking for most tissues tested, whereas β-actin was most suitable only for ovarian. Conclusions: These data reinforce the need for species-specific HKG validation and provide an appropriate panel of reference markers for gene expression studies in the M. amazonicum.
Thesagro:	Camarão Macrobrachium Amazonicum
Palavras-chave:	RT-qPCR Expressao genetica
ISSN:	2073-4425.
Digital Object Identifier:	https://doi.org/10.3390/genes17010026
Tipo do material:	Artigo de periódico
Acesso:	openAccess
Aparece nas coleções:	Artigo em periódico indexado (CNPASA)