Indian Journal of Animal Research
SCOPUSWeb of Science
  • Year: 2024
  • Volume: 58
  • Issue: 11

Identification of suitable reference genes for qPCR analysis of 4T1 mouse mammary tumor cell line

  • Author:
  • Richa Arora1,*, Waseem Akram Malla1, Arpit Tyagi2, Shikha Saxena3, Sonalika Mahajan1, Basavaraj Sajjanar1, Ashok Kumar Tiwari4
  • Total Page Count: 6
  • Page Number: 1872 to 1877

1Division of Veterinary Biotechnology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India

2GB Pant University of Agriculture and Technology, Pantnagar-263145, Uttarakhand, India

3Division of Animal Genetics, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India

4Central Avian Research Institute, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India

*Corresponding Author: Richa Arora, Division of Veterinary Biotechnology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India, Email: richaarora5521@gmail.com

Online published on 11 June, 2025.

Abstract

Identification of candidate reference genes for real time PCR study is a preliminary requirement to normalize experimental data and thus, deduce a reliable conclusion. Complex tissues like mouse mammary gland constitutes various cell types which makes it difficult to identify reference gene constantly expressing under different experimental conditions.

In this study we have identified suitable reference genes for 4T1 tumor cell line derived from mouse mammary tumor cells. We have studied four genes namely Gapdh, Actb, Prdx1 and Ctbp1 for their expression stability in CPV2.NS1 post transfected 4T1 cells by Best Keeper.

By our study, three reference genes i.e.Prdx1, Gapdh and Ctbp1 were found to be quite correlated with the BestKeeper index, but by considering all three criteria of selection by Best Keeper algorithm, Prdx1 showed minimum standard deviation and coefficient of variation and was found to be ranked at first position by BestKeeper which suggests Prdx1 to be considered as better internal control gene among all other reference genes taken in our study for qPCR based experiments in 4T1 mouse mammary tumor cell line transfected with CPV2.NS1.

Keywords

Canine parvovirus, Mammary gland tumor, Non-structural gene 1, Oncolytic viral genes, Pearson correlation coefficient, Standard deviation