Transcriptome-based Selection and Validation of Optimal House-keeping Genes for Skin Research in Goats (Capra hircus)

Abstract Background : In quantitative real-time polymerase chain reaction (qRT-PCR) experiments, accurate and reliable target gene expression results are dependent on optimal amplification of house-keeping genes (HKGs). RNA-seq technology offers a novel approach to detect new HKGs with improved stability. Goat ( Capra hircus ) is an economically important livestock species and plays an indispensable role in the world animal fiber and meat industry. Unfortunately, uniform and reliable HKGs for skin research have not been identified in goat. Therefore, this study seeks to identify a set of stable HKGs for the skin tissue of C. hircus using high-throughput sequencing technology. Results: Based on the transcriptome dataset of 39 goat skin tissue samples, 8 genes ( SRP68 , NCBP3 , RRAGA , EIF4H , CTBP2 , PTPRA , CNBP , and EEF2 ) with relatively stable expression levels were identified and selected as new candidate HKGs. Commonly used HKGs including SDHA and YWHAZ from a previous study, and 2 conventional genes ( ACTB and GAPDH ) were also examined. Four different experimental variables: (1) different development stages, (2) hair follicle cycle stages, (3) breeds, and (4) sampling sites were used for determination and validation. Four algorithms (geNorm, NormFinder, BestKeeper, and ΔCt method) and a comprehensive algorithm (ComprFinder, developed in-house) were used to assess the stability of each HKG. It was shown that NCBP3+SDHA+PTPRA were more stably expressed than previously used genes in all conditions analysis, and that this combination was effective at normalizing target gene expression. Moreover, a new algorithm for comprehensive analysis, ComprFinder, was developed and released. Conclusion: This study presents the first list of candidate HKGs for C. hircus skin tissues based on an RNA-seq dataset. We propose that the NCBP3+SDHA+PTPRA combination could be regarded as a triplet set of HKGs in skin molecular biology experiments in C. hircus and other closely related species. In addition, we also encourage researchers who perform candidate HKG evaluations and who require comprehensive analysis to adopt our new algorithm, ComprFinder.

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Transcriptome-based Selection and Validation of Optimal House-keeping Genes for Skin Research in Goats (Capra hircus)

10.21203/rs.3.rs-17424/v3 ◽

2020 ◽

Author(s):

Jipan Zhang ◽

Chengchen Deng ◽

Jialu Li ◽

Yong-Ju Zhao

Keyword(s):

Gene Expression ◽

Target Gene ◽

High Throughput Sequencing ◽

Comprehensive Analysis ◽

Skin Tissue ◽

Capra Hircus ◽

Meat Industry ◽

Rna Seq ◽

Target Gene Expression ◽

Goat Skin

Abstract Background: In Quantitative real-time polymerase chain reaction (qRT-PCR) experiments, accurate and reliable target gene expression data is dependent on optimal amplification of house-keeping genes (HKGs). RNA-seq technology offers a novel approach to detect new HKGs with improved stability. Goat (Capra hircus) is an economically important livestock species and plays an indispensable role in the world animal fiber and meat industry. Unfortunately, uniform and reliable HKGs for use in goat skin research have not been identified. Therefore, this study seeks to identify a set of stable HKGs for the skin tissue of C. hircus using high-throughput sequencing technology.Results: Based on the transcriptome dataset of 39 goat skin tissue samples, 8 genes (SRP68, NCBP3, RRAGA, EIF4H, CTBP2, PTPRA, CNBP, and EEF2) with relatively stable expression levels were identified and selected as new candidate HKGs. Commonly used HKGs including SDHA and YWHAZ from a previous study, and 2 conventional genes (ACTB and GAPDH) were also examined. Four different experimental variables: (1) different development stages, (2) hair follicle cycle stages, (3) breeds and (4) sampling sites, were used for determination and validation. Four algorithms (geNorm, NormFinder, BestKeeper, and ΔCt method) and a comprehensive algorithm (ComprFinder, developed in-house) were used to assess the stability of each HKG. It was shown that NCBP3+SDHA+PTPRA were more stably expressed than previously used genes in all conditions analyzed, and that this combination was effective at normalizing target gene expression. Moreover, a new algorithm for comprehensive analysis, ComprFinder, was developed and released.Conclusion: This study presents the first data of candidate HKGs for C. hircus skin tissues based on an RNA-seq dataset. We propose that the NCBP3+SDHA+PTPRA combination be used as a triplet set of HKGs in skin molecular biology experiments in C. hircus and other closely related species to standardize analyses across studies. In addition, we also encourage researchers who perform candidate HKG evaluations and who have the need for comprehensive analysis to adopt our new algorithm, ComprFinder.

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Transcriptome-based Selection and Validation of Optimal House-keeping Genes for Skin Research in Goats (Capra hircus)

10.21203/rs.3.rs-17424/v2 ◽

2020 ◽

Author(s):

JIPAN ZHANG ◽

Chengchen Deng ◽

Jialu Li ◽

Yong-Ju Zhao

Keyword(s):

Gene Expression ◽

Target Gene ◽

High Throughput Sequencing ◽

Comprehensive Analysis ◽

Skin Tissue ◽

Capra Hircus ◽

Meat Industry ◽

Rna Seq ◽

Target Gene Expression ◽

Goat Skin

Abstract Background: In Quantitative real-time polymerase chain reaction (qRT-PCR) experiments, accurate and reliable target gene expression data is dependent on optimal amplification of house-keeping genes (HKGs). RNA-seq technology offers a novel approach to detect new HKGs with improved stability. Goat (Capra hircus) is an economically important livestock species and plays an indispensable role in the world animal fiber and meat industry. Unfortunately, uniform and reliable HKGs for use in goat skin research have not been identified. Therefore, this study seeks to identify a set of stable HKGs for the skin tissue of C. hircus using high-throughput sequencing technology.Results: Based on the transcriptome dataset of 39 goat skin tissue samples, 8 genes (SRP68, NCBP3, RRAGA, EIF4H, CTBP2, PTPRA, CNBP, and EEF2) with relatively stable expression levels were identified and selected as new candidate HKGs. Commonly used HKGs including SDHA and YWHAZ from a previous study, and 2 conventional genes (ACTB and GAPDH) were also examined. Four different experimental variables: (1) different development stages, (2) hair follicle cycle stages, (3) breeds and (4) sampling sites, were used for determination and validation. Four algorithms (geNorm, NormFinder, BestKeeper, and ΔCt method) and a comprehensive algorithm (ComprFinder, developed in-house) were used to assess the stability of each HKG. It was shown that NCBP3+SDHA+PTPRA were more stably expressed than previously used genes in all conditions analysed, and that this combination was effective at normalizing target gene expression. Moreover, a new algorithm for comprehensive analysis, ComprFinder, was developed and released.Conclusion: This study presents the first data of candidate HKGs for C. hircus skin tissues based on an RNA-seq dataset. We propose that the NCBP3+SDHA+PTPRA combination be used as a triplet set of HKGs in skin molecular biology experiments in C. hircus and other closely related species to standardize analyses across studies. In addition, we also encourage researchers who perform candidate HKG evaluations and who have the need for comprehensive analysis to adopt our new algorithm, ComprFinder.

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Transcriptome-based Selection and Validation of Optimal House-keeping Genes for Skin Research in Goats (Capra hircus)

10.21203/rs.3.rs-17424/v4 ◽

2020 ◽

Author(s):

Jipan Zhang ◽

Chengchen Deng ◽

Jialu Li ◽

Yong-Ju Zhao

Keyword(s):

Gene Expression ◽

Target Gene ◽

High Throughput Sequencing ◽

Comprehensive Analysis ◽

Skin Tissue ◽

Capra Hircus ◽

Meat Industry ◽

Rna Seq ◽

Target Gene Expression ◽

Goat Skin

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Transcriptome-based Selection and Validation of Optimal House-keeping Genes for Skin Research in Goats (Capra hircus)

10.21203/rs.3.rs-17424/v1 ◽

2020 ◽

Author(s):

Jipan Zhang ◽

Chengchen Deng ◽

Jialu Li ◽

Yong-Ju Zhao

Keyword(s):

Gene Expression ◽

Target Gene ◽

High Throughput Sequencing ◽

Capra Hircus ◽

Meat Industry ◽

Rna Seq ◽

Target Gene Expression ◽

Novel Approach ◽

Improved Stability ◽

The Stability

Abstract Background: In Quantitative real-time polymerase chain reaction (qRT-PCR) experiments, accurate and reliable target gene expression data is dependent on optimal amplification of house-keeping genes (HKGs). The RNA-seq technology offers a novel approach to detect new HKGs with improved stability. Goat (Capra hircus) is an economically important livestock species, and plays an indispensable role in the world animal fiber and meat industry. Unfortunately, uniform and reliable HKGs be used in skin research of goats have not been identified. Therefore, this study seeks to identify a set of stable HKGs for the skin tissue of C. hircus using the new high-throughput sequencing technology.Results: Based on the transcriptome dataset of 39 goat skin tissues, 8 genes (SRP68, NCBP3, RRAGA, EIF4H, CTBP2, PTPRA, CNBP, and EEF2) with relatively stable expression levels were identified and selected as new candidate HKGs. The classical HKGs including SDHA and YWHAZ from a previous study, and 2 conventional genes (ACTB and GAPDH) were also considered. Four different experimental materials: (1) different development stages, (2) hair follicle cycle stages, (3) breeds and (4) sampling sites were provided for determination and validation. Four algorithms (geNorm, NormFinder, BestKeeper, and ΔCt method) and a comprehensive algorithm (ComprFinder, developed in-house) were used to assess the stability of each HKG. It was observed that NCBP3+SDHA+PTPRA was more stably expressed than previously used genes, in all conditions analyzed. This combination was effective at normalizing target gene expression. Moreover, a new algorithm, ComprFinder, was developed and released for comprehensive analysis.Conclusion: This study presents the first data of candidate HKGs selection for skin tissues of C. hircus based on an RNA-seq dataset. We propose the use of the NCBP3+SDHA+PTPRA combination as the triplet HKGs in skin molecular biology in C. hircus and other closely related species in order to standardize analyses across studies. In addition, we also encourage researchers who are performing candidate HKG evaluations and have the needs of a comprehensive analysis to adopt our new algorithm, ComprFinder.

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2049-P: The Chd4 Helicase of the Nucleosome Remodeling and Deacetylase Complex Modulates Pdx1 Target Gene Expression In Vitro

Diabetes ◽

10.2337/db20-2049-p ◽

2020 ◽

Vol 69 (Supplement 1) ◽

pp. 2049-P

Author(s):

REBECCA K. DAVIDSON ◽

NOLAN CASEY ◽

JASON SPAETH

Keyword(s):

Gene Expression ◽

Target Gene ◽

Nucleosome Remodeling ◽

Target Gene Expression

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On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

Applied Microbiology and Biotechnology ◽

10.1007/s00253-021-11269-z ◽

2021 ◽

Author(s):

Philipp Moritz Fricke ◽

Angelika Klemm ◽

Michael Bott ◽

Tino Polen

Keyword(s):

Gene Expression ◽

Acetic Acid ◽

Literature Search ◽

Target Gene ◽

Target Genes ◽

Recombinant Dna ◽

Acetic Acid Bacteria ◽

Homoserine Lactone ◽

Expression Systems ◽

Target Gene Expression

Abstract Acetic acid bacteria (AAB) are valuable biocatalysts for which there is growing interest in understanding their basics including physiology and biochemistry. This is accompanied by growing demands for metabolic engineering of AAB to take advantage of their properties and to improve their biomanufacturing efficiencies. Controlled expression of target genes is key to fundamental and applied microbiological research. In order to get an overview of expression systems and their applications in AAB, we carried out a comprehensive literature search using the Web of Science Core Collection database. The Acetobacteraceae family currently comprises 49 genera. We found overall 6097 publications related to one or more AAB genera since 1973, when the first successful recombinant DNA experiments in Escherichia coli have been published. The use of plasmids in AAB began in 1985 and till today was reported for only nine out of the 49 AAB genera currently described. We found at least five major expression plasmid lineages and a multitude of further expression plasmids, almost all enabling only constitutive target gene expression. Only recently, two regulatable expression systems became available for AAB, an N-acyl homoserine lactone (AHL)-inducible system for Komagataeibacter rhaeticus and an l-arabinose-inducible system for Gluconobacter oxydans. Thus, after 35 years of constitutive target gene expression in AAB, we now have the first regulatable expression systems for AAB in hand and further regulatable expression systems for AAB can be expected. Key points • Literature search revealed developments and usage of expression systems in AAB. • Only recently 2 regulatable plasmid systems became available for only 2 AAB genera. • Further regulatable expression systems for AAB are in sight.

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