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ArticleMolecular biology reports2024

Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR.

Shivali Pandita, Hera Alam, Radha Shivhare, Manisha Singh, Sanchita Singh, Geetanjali Mishra, Praveen C Verma

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Article in Molecular biology reports, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

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3 citing papers in PubMed.

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  3. 3-O-Acetyl-11-Keto-Oncology research · 2025
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5 · Who and what money

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7 authors.

Shivali PanditaMolecular Biology and Biotechnology Division, CSIR-National Botanical Research Institute, (Council of Scientific and Industrial Research) Rana Pratap Marg, Lucknow, UP, 226001, India.
Hera AlamDepartment of Zoology, University of Lucknow, Lucknow, Uttar Pradesh, 226007, India.
Radha ShivhareMicrobial Technologies Division, CSIR-National Botanical Research Institute, (Council of Scientific and Industrial Research) Rana Pratap Marg, Lucknow, UP, 226001, India.
Manisha SinghMolecular Biology and Biotechnology Division, CSIR-National Botanical Research Institute, (Council of Scientific and Industrial Research) Rana Pratap Marg, Lucknow, UP, 226001, India.
Sanchita SinghMolecular Biology and Biotechnology Division, CSIR-National Botanical Research Institute, (Council of Scientific and Industrial Research) Rana Pratap Marg, Lucknow, UP, 226001, India.
Geetanjali MishraDepartment of Zoology, University of Lucknow, Lucknow, Uttar Pradesh, 226007, India.
Praveen C VermaMolecular Biology and Biotechnology Division, CSIR-National Botanical Research Institute, (Council of Scientific and Industrial Research) Rana Pratap Marg, Lucknow, UP, 226001, India. praveencverma@nbri.res.in.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundRegeneration 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

resultsIn 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.

conclusionAccording 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.

Indexed as

ColeopteraGene Expression ProfilingGenes, EssentialReal-Time Polymerase Chain ReactionRegenerationAnimalsGenes, InsectReference StandardsTranscriptomeCheilomenes sexmaculataGeNormNormFinderqRT-PCRReference genesRefFinderRegeneration

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