Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR
Background Regeneration is a fascinating phenomenon that has intrigued scientists for a long time. Cheilomenes sexmaculata (Fabricius), a zig-zag ladybird beetle, possesses a high capacity for limb regeneration. The molecular mechanics of the zig-zag ladybird beetle are under-explored. Current resea...
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| description | Background
Regeneration is a fascinating phenomenon that has intrigued scientists for a long time.
Cheilomenes sexmaculata
(Fabricius), a zig-zag ladybird beetle, possesses a high capacity for limb regeneration. The molecular mechanics of the zig-zag ladybird beetle are under-explored. Current research trends are focused on uncovering functional genes associated with limb regeneration. Most of these investigations involve quantitative real-time PCR (qRT-PCR) for their rapid and accurate analysis of gene expression levels. Hence, a stable and suitable reference gene is required to normalize the gene expression data.
Methods and results
In this study, five housekeeping genes were selected from the transcriptomics data (
in-house
unpublished data) of
C. sexmaculata
(Fabricius). The expression stabilities of the selected genes were evaluated under different time intervals post-amputation using geNorm, normFinder, and refFinder software.
Actin
was revealed to be the most stable housekeeping gene, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase.
A target gene named
engrailed
(an important segment-forming gene) was used to validate the selected reference genes. The expression levels were found to be consistent with the transcriptomics results.
Conclusion
According to our study,
actin
, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase
, serve as the most stable reference genes and are suitable for regeneration-related research. This study is a groundbreaking effort to identify the most stable reference gene for limb regeneration in
C. sexmaculata
(Fabricius), and the findings can be applied to other related insect species. |
| doi_str_mv | 10.1007/s11033-024-10062-1 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_3154169754</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>3154169754</sourcerecordid><originalsourceid>FETCH-LOGICAL-c289t-827a8881a4b8e5bb1d826909dfaeaf611c2c6e407dad67a9990095eb20a8622d3</originalsourceid><addsrcrecordid>eNqNkU-LFDEQxYMo7rj6BTxIwIuXaCXpTidHGfwHC8q6npvqTvWapf_MJN3Lzpfxs5qdHhU8iKfiUb_3Cuox9lzCawlQvUlSgtYCVCGyNkrIB2wjy0qLwlX2IduABikKW8oz9iSlGwAoZFU-ZmfaFa7UldmwH1-pp3YO08hx9PwW--DxKKeOR-oo0tgSv6aREu-myPcLjnOYM3NLnO52kVJa3dgfUkir756PxxwRqceZ_CkijHz7nUI_DUeZ6G7AdskE8uaQjdiLOQzE95dX4sv28il71GGf6NlpnrNv799dbT-Ki88fPm3fXohWWTcLqyq01kosGktl00hvlXHgfIeEnZGyVa2hAiqP3lTonANwJTUK0BqlvD5nr9bcXZz2C6W5HkJqqe9xpGlJtZZlIY2ryuI_UKUtGKtURl_-hd5MS8yPWimnrDaQKbVSbZxSyj-vdzEMGA-1hPq-6Hotus5F18eia5lNL07RSzOQ_2351WwG9AqkvBqvKf65_Y_Yn_lRtWM</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>3123928360</pqid></control><display><type>article</type><title>Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR</title><source>MEDLINE</source><source>SpringerLink (Online service)</source><creator>Pandita, Shivali ; Alam, Hera ; Shivhare, Radha ; Singh, Manisha ; Singh, Sanchita ; Mishra, Geetanjali ; Verma, Praveen C.</creator><creatorcontrib>Pandita, Shivali ; Alam, Hera ; Shivhare, Radha ; Singh, Manisha ; Singh, Sanchita ; Mishra, Geetanjali ; Verma, Praveen C.</creatorcontrib><description>Background
Regeneration is a fascinating phenomenon that has intrigued scientists for a long time.
Cheilomenes sexmaculata
(Fabricius), a zig-zag ladybird beetle, possesses a high capacity for limb regeneration. The molecular mechanics of the zig-zag ladybird beetle are under-explored. Current research trends are focused on uncovering functional genes associated with limb regeneration. Most of these investigations involve quantitative real-time PCR (qRT-PCR) for their rapid and accurate analysis of gene expression levels. Hence, a stable and suitable reference gene is required to normalize the gene expression data.
Methods and results
In this study, five housekeeping genes were selected from the transcriptomics data (
in-house
unpublished data) of
C. sexmaculata
(Fabricius). The expression stabilities of the selected genes were evaluated under different time intervals post-amputation using geNorm, normFinder, and refFinder software.
Actin
was revealed to be the most stable housekeeping gene, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase.
A target gene named
engrailed
(an important segment-forming gene) was used to validate the selected reference genes. The expression levels were found to be consistent with the transcriptomics results.
Conclusion
According to our study,
actin
, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase
, serve as the most stable reference genes and are suitable for regeneration-related research. This study is a groundbreaking effort to identify the most stable reference gene for limb regeneration in
C. sexmaculata
(Fabricius), and the findings can be applied to other related insect species.</description><identifier>ISSN: 0301-4851</identifier><identifier>ISSN: 1573-4978</identifier><identifier>EISSN: 1573-4978</identifier><identifier>DOI: 10.1007/s11033-024-10062-1</identifier><identifier>PMID: 39495376</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Actin ; Amputation ; Animal Anatomy ; Animal Biochemistry ; Animals ; Biomedical and Life Sciences ; Cheilomenes ; Cheilomenes sexmaculata ; Coleoptera - genetics ; Coleoptera - physiology ; computer software ; Dehydrogenases ; Elongation ; Gene expression ; Gene Expression Profiling - methods ; Gene Expression Profiling - standards ; genes ; Genes, Essential - genetics ; Genes, Insect - genetics ; Glyceraldehyde-3-phosphate dehydrogenase ; Histology ; insects ; Life Sciences ; mechanics ; Morphology ; Original Article ; peptide elongation factors ; quantitative polymerase chain reaction ; Real-Time Polymerase Chain Reaction - methods ; Real-Time Polymerase Chain Reaction - standards ; Reference Standards ; Regeneration ; Regeneration - genetics ; species ; Transcriptome - genetics ; Transcriptomics</subject><ispartof>Molecular biology reports, 2024-12, Vol.51 (1), p.1118-1118, Article 1118</ispartof><rights>The Author(s), under exclusive licence to Springer Nature B.V. 2024 Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</rights><rights>2024. The Author(s), under exclusive licence to Springer Nature B.V.</rights><rights>Copyright Springer Nature B.V. Dec 2024</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c289t-827a8881a4b8e5bb1d826909dfaeaf611c2c6e407dad67a9990095eb20a8622d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11033-024-10062-1$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11033-024-10062-1$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/39495376$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pandita, Shivali</creatorcontrib><creatorcontrib>Alam, Hera</creatorcontrib><creatorcontrib>Shivhare, Radha</creatorcontrib><creatorcontrib>Singh, Manisha</creatorcontrib><creatorcontrib>Singh, Sanchita</creatorcontrib><creatorcontrib>Mishra, Geetanjali</creatorcontrib><creatorcontrib>Verma, Praveen C.</creatorcontrib><title>Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR</title><title>Molecular biology reports</title><addtitle>Mol Biol Rep</addtitle><addtitle>Mol Biol Rep</addtitle><description>Background
Regeneration is a fascinating phenomenon that has intrigued scientists for a long time.
Cheilomenes sexmaculata
(Fabricius), a zig-zag ladybird beetle, possesses a high capacity for limb regeneration. The molecular mechanics of the zig-zag ladybird beetle are under-explored. Current research trends are focused on uncovering functional genes associated with limb regeneration. Most of these investigations involve quantitative real-time PCR (qRT-PCR) for their rapid and accurate analysis of gene expression levels. Hence, a stable and suitable reference gene is required to normalize the gene expression data.
Methods and results
In this study, five housekeeping genes were selected from the transcriptomics data (
in-house
unpublished data) of
C. sexmaculata
(Fabricius). The expression stabilities of the selected genes were evaluated under different time intervals post-amputation using geNorm, normFinder, and refFinder software.
Actin
was revealed to be the most stable housekeeping gene, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase.
A target gene named
engrailed
(an important segment-forming gene) was used to validate the selected reference genes. The expression levels were found to be consistent with the transcriptomics results.
Conclusion
According to our study,
actin
, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase
, serve as the most stable reference genes and are suitable for regeneration-related research. This study is a groundbreaking effort to identify the most stable reference gene for limb regeneration in
C. sexmaculata
(Fabricius), and the findings can be applied to other related insect species.</description><subject>Actin</subject><subject>Amputation</subject><subject>Animal Anatomy</subject><subject>Animal Biochemistry</subject><subject>Animals</subject><subject>Biomedical and Life Sciences</subject><subject>Cheilomenes</subject><subject>Cheilomenes sexmaculata</subject><subject>Coleoptera - genetics</subject><subject>Coleoptera - physiology</subject><subject>computer software</subject><subject>Dehydrogenases</subject><subject>Elongation</subject><subject>Gene expression</subject><subject>Gene Expression Profiling - methods</subject><subject>Gene Expression Profiling - standards</subject><subject>genes</subject><subject>Genes, Essential - genetics</subject><subject>Genes, Insect - genetics</subject><subject>Glyceraldehyde-3-phosphate dehydrogenase</subject><subject>Histology</subject><subject>insects</subject><subject>Life Sciences</subject><subject>mechanics</subject><subject>Morphology</subject><subject>Original Article</subject><subject>peptide elongation factors</subject><subject>quantitative polymerase chain reaction</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Real-Time Polymerase Chain Reaction - standards</subject><subject>Reference Standards</subject><subject>Regeneration</subject><subject>Regeneration - genetics</subject><subject>species</subject><subject>Transcriptome - genetics</subject><subject>Transcriptomics</subject><issn>0301-4851</issn><issn>1573-4978</issn><issn>1573-4978</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU-LFDEQxYMo7rj6BTxIwIuXaCXpTidHGfwHC8q6npvqTvWapf_MJN3Lzpfxs5qdHhU8iKfiUb_3Cuox9lzCawlQvUlSgtYCVCGyNkrIB2wjy0qLwlX2IduABikKW8oz9iSlGwAoZFU-ZmfaFa7UldmwH1-pp3YO08hx9PwW--DxKKeOR-oo0tgSv6aREu-myPcLjnOYM3NLnO52kVJa3dgfUkir756PxxwRqceZ_CkijHz7nUI_DUeZ6G7AdskE8uaQjdiLOQzE95dX4sv28il71GGf6NlpnrNv799dbT-Ki88fPm3fXohWWTcLqyq01kosGktl00hvlXHgfIeEnZGyVa2hAiqP3lTonANwJTUK0BqlvD5nr9bcXZz2C6W5HkJqqe9xpGlJtZZlIY2ryuI_UKUtGKtURl_-hd5MS8yPWimnrDaQKbVSbZxSyj-vdzEMGA-1hPq-6Hotus5F18eia5lNL07RSzOQ_2351WwG9AqkvBqvKf65_Y_Yn_lRtWM</recordid><startdate>20241201</startdate><enddate>20241201</enddate><creator>Pandita, Shivali</creator><creator>Alam, Hera</creator><creator>Shivhare, Radha</creator><creator>Singh, Manisha</creator><creator>Singh, Sanchita</creator><creator>Mishra, Geetanjali</creator><creator>Verma, Praveen C.</creator><general>Springer Netherlands</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TK</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>K9.</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope></search><sort><creationdate>20241201</creationdate><title>Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR</title><author>Pandita, Shivali ; Alam, Hera ; Shivhare, Radha ; Singh, Manisha ; Singh, Sanchita ; Mishra, Geetanjali ; Verma, Praveen C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c289t-827a8881a4b8e5bb1d826909dfaeaf611c2c6e407dad67a9990095eb20a8622d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Actin</topic><topic>Amputation</topic><topic>Animal Anatomy</topic><topic>Animal Biochemistry</topic><topic>Animals</topic><topic>Biomedical and Life Sciences</topic><topic>Cheilomenes</topic><topic>Cheilomenes sexmaculata</topic><topic>Coleoptera - genetics</topic><topic>Coleoptera - physiology</topic><topic>computer software</topic><topic>Dehydrogenases</topic><topic>Elongation</topic><topic>Gene expression</topic><topic>Gene Expression Profiling - methods</topic><topic>Gene Expression Profiling - standards</topic><topic>genes</topic><topic>Genes, Essential - genetics</topic><topic>Genes, Insect - genetics</topic><topic>Glyceraldehyde-3-phosphate dehydrogenase</topic><topic>Histology</topic><topic>insects</topic><topic>Life Sciences</topic><topic>mechanics</topic><topic>Morphology</topic><topic>Original Article</topic><topic>peptide elongation factors</topic><topic>quantitative polymerase chain reaction</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Real-Time Polymerase Chain Reaction - standards</topic><topic>Reference Standards</topic><topic>Regeneration</topic><topic>Regeneration - genetics</topic><topic>species</topic><topic>Transcriptome - genetics</topic><topic>Transcriptomics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pandita, Shivali</creatorcontrib><creatorcontrib>Alam, Hera</creatorcontrib><creatorcontrib>Shivhare, Radha</creatorcontrib><creatorcontrib>Singh, Manisha</creatorcontrib><creatorcontrib>Singh, Sanchita</creatorcontrib><creatorcontrib>Mishra, Geetanjali</creatorcontrib><creatorcontrib>Verma, Praveen C.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><jtitle>Molecular biology reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pandita, Shivali</au><au>Alam, Hera</au><au>Shivhare, Radha</au><au>Singh, Manisha</au><au>Singh, Sanchita</au><au>Mishra, Geetanjali</au><au>Verma, Praveen C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR</atitle><jtitle>Molecular biology reports</jtitle><stitle>Mol Biol Rep</stitle><addtitle>Mol Biol Rep</addtitle><date>2024-12-01</date><risdate>2024</risdate><volume>51</volume><issue>1</issue><spage>1118</spage><epage>1118</epage><pages>1118-1118</pages><artnum>1118</artnum><issn>0301-4851</issn><issn>1573-4978</issn><eissn>1573-4978</eissn><abstract>Background
Regeneration is a fascinating phenomenon that has intrigued scientists for a long time.
Cheilomenes sexmaculata
(Fabricius), a zig-zag ladybird beetle, possesses a high capacity for limb regeneration. The molecular mechanics of the zig-zag ladybird beetle are under-explored. Current research trends are focused on uncovering functional genes associated with limb regeneration. Most of these investigations involve quantitative real-time PCR (qRT-PCR) for their rapid and accurate analysis of gene expression levels. Hence, a stable and suitable reference gene is required to normalize the gene expression data.
Methods and results
In this study, five housekeeping genes were selected from the transcriptomics data (
in-house
unpublished data) of
C. sexmaculata
(Fabricius). The expression stabilities of the selected genes were evaluated under different time intervals post-amputation using geNorm, normFinder, and refFinder software.
Actin
was revealed to be the most stable housekeeping gene, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase.
A target gene named
engrailed
(an important segment-forming gene) was used to validate the selected reference genes. The expression levels were found to be consistent with the transcriptomics results.
Conclusion
According to our study,
actin
, along with
elongation factor 2
and
glyceraldehyde-3-phosphate dehydrogenase
, serve as the most stable reference genes and are suitable for regeneration-related research. This study is a groundbreaking effort to identify the most stable reference gene for limb regeneration in
C. sexmaculata
(Fabricius), and the findings can be applied to other related insect species.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>39495376</pmid><doi>10.1007/s11033-024-10062-1</doi><tpages>1</tpages></addata></record> |
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| subjects | Actin Amputation Animal Anatomy Animal Biochemistry Animals Biomedical and Life Sciences Cheilomenes Cheilomenes sexmaculata Coleoptera - genetics Coleoptera - physiology computer software Dehydrogenases Elongation Gene expression Gene Expression Profiling - methods Gene Expression Profiling - standards genes Genes, Essential - genetics Genes, Insect - genetics Glyceraldehyde-3-phosphate dehydrogenase Histology insects Life Sciences mechanics Morphology Original Article peptide elongation factors quantitative polymerase chain reaction Real-Time Polymerase Chain Reaction - methods Real-Time Polymerase Chain Reaction - standards Reference Standards Regeneration Regeneration - genetics species Transcriptome - genetics Transcriptomics |
| title | Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR |
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