keyword
https://read.qxmd.com/read/38367736/selection-identification-and-evaluation-of-optimal-reference-genes-in-chinese-sturgeon-acipenser-sinensis-under-polypropylene-microplastics-stress
#1
JOURNAL ARTICLE
Xu Cheng, Kan Xiao, Wei Jiang, Guangyuan Peng, Pei Chen, Tingting Shu, Hongtao Huang, Xuetao Shi, Jing Yang
Polypropylene microplastics (PP-MPs) are emerging environmental contaminants that have the potential to cause adverse effects on aquatic organisms. Reverse transcriptase quantitative real-time polymerase chain reaction (RT-qPCR) is a valuable tool for assessing the gene expression profiles under PP-MPs stress. To obtain an accurate gene expression profile of tissue inflammation and apoptosis that reflects the molecular mechanisms underlying the impact of PP-MPs on Chinese sturgeon, identifying reliable reference genes is crucial for RT-qPCR analysis...
February 15, 2024: Science of the Total Environment
https://read.qxmd.com/read/38302649/comprehensive-evaluation-and-validation-of-optimal-reference-genes-for-normalization-of-qpcr-data-in-different-caprine-tissues
#2
JOURNAL ARTICLE
Sonika Ahlawat, Mahanthi Vasu, Vikas Choudhary, Reena Arora, Rekha Sharma, M A Mir, Manoj Kumar Singh
BACKGROUND: Quantitative real-time PCR (qPCR) is a highly reliable method for validating gene expression data in molecular studies due to its sensitivity, specificity, and efficiency. To ensure accurate qPCR results, it's essential to normalize the expression data using stable reference genes. METHODS: This study aimed to identify suitable reference genes for qPCR studies in goats by evaluating 18 candidate reference genes (ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT, and YWHAZ) in 10 different caprine tissues (heart, intestine, kidney, liver, lung, muscle, rumen, skin, spleen, and testis)...
February 1, 2024: Molecular Biology Reports
https://read.qxmd.com/read/38092161/identification-and-validation-of-stable-reference-genes-for-expression-profiling-of-target-genes-in-diverse-ovine-tissues
#3
JOURNAL ARTICLE
Mahanthi Vasu, Sonika Ahlawat, Vikas Choudhary, Rashmeet Kaur, Reena Arora, Rekha Sharma, Upasna Sharma, Pooja Chhabra, M A Mir, Manoj Kumar Singh
Quantitative PCR (qPCR) is a widely-used technique for quantifying the expression of target genes across various tissues, as well as under different pathological and physiological conditions. One of the challenges associated with this method is the need to identify optimal reference genes (RGs) that maintain consistent expression levels under diverse experimental settings, thereby ensuring accurate biological interpretation. In this study, we conducted a thorough analysis of 18 candidate RGs (ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT, and YWHAZ) across 10 ovine tissues (muscle, skin, kidney, liver, intestine, rumen, lung, testis, heart, and spleen) obtained from five individual sheep...
December 11, 2023: Gene
https://read.qxmd.com/read/36961042/systematic-identification-of-the-optimal-housekeeping-genes-for-accurate-transcriptomic-and-proteomic-profiling-of-tissues-following-complex-traumatic-injury
#4
JOURNAL ARTICLE
Andrea H Dragon, Cassie J Rowe, Alisha M Rhodes, Olivia L Pak, Thomas A Davis, Elsa Ronzier
Trauma triggers critical molecular and cellular signaling cascades that drive biological outcomes and recovery. Variations in the gene expression of common endogenous reference housekeeping genes (HKGs) used in data normalization differ between tissue types and pathological states. Systematically, we investigated the gene stability of nine HKGs ( Actb , B2m , Gapdh , Hprt1 , Pgk1 , Rplp0 , Rplp2 , Tbp , and Tfrc ) from tissues prone to remote organ dysfunction (lung, liver, kidney, and muscle) following extremity trauma...
February 23, 2023: Methods and Protocols
https://read.qxmd.com/read/36696727/identification-of-reliable-reference-genes-for-gene-expression-studies-in-mouse-models-under-microplastics-stress
#5
JOURNAL ARTICLE
Ju Mu, Yu Wang, Miao Wang, Dayi Zhang, Mingying Liu
Microplastics (MPs) and nanoplastics (NPs) have been deemed to be newly emerged contaminants interfering with various physiological processes closely related with gene expression alteration. Reverse transcriptase quantitative real-time polymerase chain reaction (RT-qPCR) serves as a powerful tool to assess gene expression, however highly dependent on a reliable reference gene. Therefore, it is necessary to identify stable reference genes for gene expression study under MP or NP stress. We constructed a mouse model postexposure to polypropylene microplastics (PP-MPs) to assess PP-MPs bioaccumulation in kidney, evaluate the kidney pathological changes, and then explore potential reference genes via RT-qPCR...
January 23, 2023: Ecotoxicology and Environmental Safety
https://read.qxmd.com/read/36467654/transcriptome-based-selection-and-validation-of-optimal-reference-genes-in-perirenal-adipose-developing-of-goat-capra-hircus
#6
JOURNAL ARTICLE
Le Zhao, Haili Yang, Xingchun Li, Yumei Zhou, Taolu Liu, Yongju Zhao
Brown adipose tissue (BAT) is mainly present in young mammals and is important for maintaining body temperature in neonatal mammals because of its ability to produce non-shivering thermogenesis. There is usually a large amount of BAT around the kidneys of newborn kids, but the BAT gradually "whiting" after birth. Screening and validating appropriate reference genes is a prerequisite for further studying the mechanism of goat brown adipose tissue "whiting" during the early stages. In this study, the expression stability of 17 candidate reference genes: 12 COPS8, SAP18, IGF2R, PARL, SNRNP200, ACTG1, CLTA, GANAB, GABARAP, PCBP2, CTSB , and CD151 ) selected based on previous transcriptome data as new candidate reference genes, 3 ( PFDN5, CTNNB1 , and EIF3M ) recommended in previous studies, and 2 traditional reference genes ( ACTB and GAPDH ) was evaluated...
2022: Frontiers in Veterinary Science
https://read.qxmd.com/read/36222316/evaluation-of-housekeeping-genes-as-references-for-quantitative-real-time-pcr-analysis-of-european-eel-anguilla-anguilla
#7
JOURNAL ARTICLE
Ying-Ying Li, Xi Chen, Jin-Xian Yang, Qiang Chen, Tie-Ying Song, Jun-Qing Ge
Anguilla spp. is an important aquaculture species for which an increasing number of reference genes are being identified and applied. In this study, five housekeeping genes (RPL7, 18S, EF1A, ACTB, and GAPDH) were chosen to evaluate their reliability as reference genes for quantitative real-time PCR (qPCR) for the study of Anguilla anguilla. The expression of the selected genes in different eel tissues was determined by qPCR at different growth stages or upon challenge by Anguillid herpesvirus (AngHV), and the expression levels of these genes were then compared and evaluated with the geNorm and NormFinder algorithms...
October 12, 2022: Journal of Fish Biology
https://read.qxmd.com/read/35962323/identification-and-validation-of-stable-reference-genes-for-quantitative-real-time-pcr-in-different-minipig-tissues-at-developmental-stages
#8
JOURNAL ARTICLE
Jeongah Song, Jeonghee Cho, Jeongsik Park, Jeong Ho Hwang
BACKGROUND: Quantitative real time PCR (qPCR) is a powerful tool to evaluate mRNA expression level. However, reliable qPCR results require normalization with validated reference gene(s). In this study, we investigated stable reference genes in seven tissues according to four developmental stages in minipigs. Six candidate reference genes and one target gene (ACE2) were selected and qPCR was performed. BestKeeper, geNorm, NormFinder, and delta Ct method through the RefFinder web-based tool were used to evaluate the stability of candidate reference genes...
August 13, 2022: BMC Genomics
https://read.qxmd.com/read/35753622/relative-expression-and-validation-of-aeromonas-salmonicida-subsp-salmonicida-reference-genes-during-ex-vivo-and-in-vivo-fish-infection
#9
JOURNAL ARTICLE
Manuel Soto-Dávila, Setu Chakraborty, Javier Santander
The genus Aeromonas is found worldwide in freshwater and marine environments and has been implicated in the etiology of human and animal diseases. In fish, among Aeromonas species, A. salmonicida causes massive mortality and great economic losses in marine and continental aquaculture species. Currently, several aspects of the clinical signs and pathogenesis of this Gram-negative bacterium have been described; however, determination of an appropriate reference gene is essential to normalize cellular mRNA data remain unknown...
September 2022: Infection, Genetics and Evolution
https://read.qxmd.com/read/34861601/identification-of-best-housekeeping-genes-for-the-normalization-of-rt-qpcr-in-human-cell-lines
#10
JOURNAL ARTICLE
Leticia da Conceição Braga, Bryan Ôrtero Perez Gonçalves, Pollyanne Lacerda Coelho, Agnaldo Lopes da Silva Filho, Luciana Maria Silva
The identification of the best reference gene is a critical step to evaluate the relative change in mRNA expression of a target gene by RT-qPCR. In this work, we evaluated nineteen genes of different functional classes using Real Time Human Reference Gene Panel (Roche Applied Sciences), to identify the internal housekeeping genes (HKGs) most suitable for gene expression normalization data in human cell lines. Normal cell lines CCD-19LU (lung fibroblast), HEK-293 (epithelial cell of embryonic kidney), WI-26 VA4 (lung fibroblast), and human cancer cells, BT-549 (breast cancer), Hs 578T (breast cancer), MACL-1 (breast cancer), HeLa (cervical carcinoma), U-87 MG (glioblastoma/astrocytoma), RKO-AS45-1 (colorectal carcinoma), and TOV-21G (ovarian adenocarcinoma) were cultivated according to manufacturer's protocol...
January 2022: Acta Histochemica
https://read.qxmd.com/read/34819534/evaluation-of-endogenous-mirna-reference-genes-across-different-zebrafish-strains-developmental-stages-and-kidney-disease-models
#11
COMPARATIVE STUDY
Florian Siegerist, Tim Lange, Anna Iervolino, Thor Magnus Koppe, Weibin Zhou, Giovambattista Capasso, Karlhans Endlich, Nicole Endlich
The majority of kidney diseases arise from the loss of podocytes and from morphological changes of their highly complex foot process architecture, which inevitably leads to a reduced kidney filtration and total loss of kidney function. It could have been shown that microRNAs (miRs) play a pivotal role in the pathogenesis of podocyte-associated kidney diseases. Due to their fully functioning pronephric kidney, larval zebrafish have become a popular vertebrate model, to study kidney diseases in vivo. Unfortunately, there is no consensus about a proper normalization strategy of RT-qPCR-based miRNA expression data in zebrafish...
November 24, 2021: Scientific Reports
https://read.qxmd.com/read/34800657/characterization-of-reference-genes-for-qrt-pcr-normalization-in-rice-field-eel-monopterus-albus-to-assess-differences-in-embryonic-developmental-stages-the-early-development-of-immune-organs-and-cells-infected-with-rhabdovirus
#12
JOURNAL ARTICLE
Yuchen Liu, Shuwang Wu, Nan Jiang, Wenzhi Liu, Yong Zhou, Lingbing Zeng, Qiwang Zhong, Zhong Li, Yuding Fan
Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) has become a popular technique to assess gene expression. Suitable reference genes are normally identified first to ensure accurate normalization. The aim of the present study was to select the most stable genes in embryonic developmental stages, the early development of immune organs, and cells infected with Chinese rice-field eel rhabdovirus (CrERV) of the rice-field eel (Monopterus albus). Four reference genes, including those encoding 18S ribosomal RNA (18SrRNA), beta actin (β-actin), elongation factor 1 alpha (EF1ɑ), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were assessed using geNorm, NormFinder, BestKeeper, and RefFinder software...
January 2022: Fish & Shellfish Immunology
https://read.qxmd.com/read/34615843/reference-gene-screening-for-analyzing-gene-expression-in-the-heart-liver-spleen-lung-and-kidney-of-forest-musk-deer
#13
JOURNAL ARTICLE
Xi Wu, Wei Yang, Ziwei Ren, Jianguo Cheng, Yan Luo, Yin Wang, Zexiao Yang, Xueping Yao, Wei Zhao, Yimeng Li, Kexin Tang
The screening of reference genes for real-time quantitative PCR (qPCR) in forest musk deer (FMD) tissue is of great significance to the basic research on FMD. However, there are few reports on the stability analysis of FMD reference genes so far. In this study, We used qPCR to detect the expression levels of 11 reference gene candidates (18S rRNA, beta-actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box-binding protein [TBP], hypoxanthine phosphoribosyltransferase 1 [HPRT1], tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide [YWHAZ], hydroxymethylbilane synthase [HMBS], eukaryotic translation elongation factor 1 alpha 1 [EEF1A1], succinate dehydrogenase complex flavoprotein subunit A [SDHA], peptidylprolyl isomerase B [PPIB], and ubiquitin C [UBC]) in heart, liver, spleen, lung and kidney of FMD...
November 16, 2021: Journal of Veterinary Medical Science
https://read.qxmd.com/read/34242222/reference-genes-for-mesangial-cell-and-podocyte-qpcr-gene-expression-studies-under-high-glucose-and-renin-angiotensin-system-blocker-conditions
#14
JOURNAL ARTICLE
Nicole Dittrich Hosni, Ana Carolina Anauate, Mirian Aparecida Boim
BACKGROUND: Real-time PCR remains currently the gold standard method for gene expression studies. Identification of the best reference gene is a key point in performing high-quality qPCR, providing strong support for results, and performing as a source of bias when inappropriately chosen. Mesangial cells and podocytes, as essential cell lines to study diabetic kidney disease (DKD) physiopathology, demand accurate analysis of the reference genes used thus far to enhance the validity of gene expression studies, especially regarding high glucose (HG) and DKD treatments, with angiotensin II receptor blockers (e...
2021: PloS One
https://read.qxmd.com/read/34015681/evaluation-of-candidate-reference-genes-for-quantitative-rtqpcr-analysis-in-goldfish-carassius-auratus-l-in-healthy-and-cyhv-2-infected-fish
#15
JOURNAL ARTICLE
Arathi Dharmaratnam, Arun Sudhagar, Sundar Raj Nithianantham, Sweta Das, Thangaraj Raja Swaminathan
The accuracy of quantitative real time PCR (RTqPCR) can be attained only when a suitable reference gene is used. The gene expression for a particular gene may vary within different cells at different conditions. Hence, the suitability and stability of various potential reference genes have to be determined for expression studies. In this study, we have examined the potential of four different reference genes including β-Actin (ACTB), 18S ribosomal RNA (18S), glyceraldehyde-3P-dehydrogenase (GAPDH), and elongation factor 1 alpha (EF1AA ) in seven different tissues including gill, liver, kidney, spleen, heart, muscle and intestine of goldfish (Carassius auratus)...
July 2021: Veterinary Immunology and Immunopathology
https://read.qxmd.com/read/33178342/selection-and-validation-of-optimal-endogenous-reference-genes-for-analysis-of-quantitative-pcr-in-four-tissues-pathologically-associated-with-kidney-yang-deficiency-syndrome-following-influenza-a-infection
#16
JOURNAL ARTICLE
Yepei Fu, Jia Yang, Shanshan Fan, Shaozhe Zhao, Ruikun Du, Syed Muhammad Ali Shah, Muhammad Akram, Rong Rong, Yong Yang
Kidney-yang deficiency syndrome (KYDS) infected with the influenza virus is a suitable model to imitate a population at high-risk to influenza infection with a high rate of morbidity and mortality. However, the specific molecular mechanisms underlying this disease remain unclear. A stable reference gene is essential as an internal control for molecular biology research of this condition. Reverse-transcription-quantitative PCR (RT-qPCR) is considered an extremely sensitive technique used for absolute and relative quantification of target genes transcript levels...
December 2020: Experimental and Therapeutic Medicine
https://read.qxmd.com/read/32420381/a-novel-strategy-facilitates-reference-gene-selection-by-rt-qpcr-analysis-in-kidney-yang-deficiency-syndrome-mice-infected-with-the-influenza-a-h1n1-virus
#17
JOURNAL ARTICLE
Yepei Fu, Jia Yang, Shanshan Fan, Shaozhe Zhao, Syed Muhammad Ali Shah, Muhammad Akram, Rong Rong, Yong Yang
In reverse transcription-quantitative polymerase chain reaction (RT-qPCR) studies, endogenous reference genes are routinely used to normalize the expression of target gene studies. In order to precisely evaluate the relative expression of genes in the cells of mice suffering from Kidney Yang Deficiency Syndrome (KYDS) in response to influenza A virus (IAV) H1N1 using RT-qPCR, it is crucial to identify reliable reference genes. In the present study, 15 candidate reference genes (Actb, β 2m, Gapdh, Gusb, Tuba, Grcc10, Eif4h, Rnf187, Nedd8, Ywhae, 18S rRNA, Rpl13, Ubc, Rpl32, and Ppia) were investigated in lung cells from KYDS mice infected with IAV H1N1...
2020: BioMed Research International
https://read.qxmd.com/read/31937827/identification-of-housekeeping-genes-for-microrna-expression-analysis-in-kidney-tissues-of-pkd1-deficient-mouse-models
#18
JOURNAL ARTICLE
J J Muñoz, A C Anauate, A G Amaral, F M Ferreira, R Meca, M S Ormanji, M A Boim, L F Onuchic, I P Heilberg
Polycystic kidney disease is a complex clinical entity which comprises a group of genetic diseases that leads to renal cyst development. We evaluated the most suitable housekeeping genes for microRNA expression by RT-qPCR analyses of kidney tissues in Pkd1-deficient mouse models from a panel of five candidates genes (miR-20a, miR-25, miR-26a, miR-191 and U6) and 3 target genes (miR-17, miR-21 and let-7a) using samples from kidneys of cystic mice (Pkd1flox/flox :Nestincre , CY), non-cystic controls (Pkd1flox/flox , NC), Pkd1-haploinsufficient (Pkd1+/- , HT), wild-type controls (Pkd1+/+ , WT), severely cystic mice (Pkd1V/V , SC), wild-type controls (CO)...
January 14, 2020: Scientific Reports
https://read.qxmd.com/read/31935552/transcriptome-based-evaluation-and-validation-of-suitable-housekeeping-gene-for-quantification-real-time-pcr-under-specific-experiment-condition-in-teleost-fishes
#19
JOURNAL ARTICLE
Yunkun Li, Jiabei Han, Jiayu Wu, Dong Li, Xixi Yang, Anqi Huang, Guixian Bu, Fengyan Meng, Fanli Kong, Xiaohan Cao, Xingfa Han, Xiaofu Pan, Shiyong Yang, Xianyin Zeng, Xiaogang Du
Quantification real-time PCR (qRT-PCR) is a common method in analysis of gene expression, but the stable reference genes for the normalization analysis have not been appreciated before identifying expression pattern of genes in teleost fishes. In this study, we selected eight candidate reference genes (18S, Actin, EF-1α, 40S, B2M, TUBA, UBCE and GAPDH) basing on transcriptome analysis and the traditional housekeeping genes, and analyzed the stability of the reference genes in spleen, head kidney and head kidney leukocytes (HKL) after pathogen challenge in Schizothorax prenanti (S...
March 2020: Fish & Shellfish Immunology
https://read.qxmd.com/read/31333044/reference-gene-validation-for-quantitative-real-time-pcr-studies-in-amphibian-kidney-derived-a6-epithelial-cells
#20
JOURNAL ARTICLE
Elin Verbrugghe, An Martel, Frank Pasmans
Quantitative real-time polymerase chain reaction is a widely used technique that relies on reference genes for the normalisation of gene expression. These reference genes are constitutively expressed and must remain stable across all samples and treatments. Stability of housekeeping genes may vary and must be optimised for a specific tissue, sample or cell line. Here we present a study screening for possible reference gene candidates, eef1a1 , rpl8 , sub1.L , clta , H4 and odc1 , in the Xenopus laevis (A6) kidney cell line...
May 2019: Alternatives to Laboratory Animals: ATLA
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