The beta-actin gene promoter of rohu carp (Labeo rohita) drives reporter gene expressions in transgenic rohu and various cell lines, including spermatogonial stem cells

• Published in: Cellular & molecular biology letters Vol. 20; no. 2; pp. 237 - 247
• Main Authors: Barman, Hirak Kumar, Mohanta, Ramya, Patra, Swagat Kumar, Chakrapani, Vemulawada, Panda, Rudra Prasanna, Nayak, Swapnarani, Jena, Sasmita, Jayasankar, Pallipuram, Nandanpawar, Priyanka
• Format: Journal Article
• Language: English
• Published: England De Gruyter Open 01.06.2015
• Subjects: Actins - genetics; Animals; Animals, Genetically Modified; Cell Line; Cells, Cultured; Cyprinidae - genetics; Fish Proteins - genetics; Gene Expression; Genes, Reporter; GFP; HEK293; Humans; Insecta - genetics; K562; Labeo rohita; Male; Promoter Regions, Genetic; Rohu; SF21; Spermatogonia - cytology; Spermatogonial cells; Stem Cells - metabolism; Transfection protocol; Transgenic; β-actin promoter
• ISSN: 1425-8153; 1689-1392
• DOI: 10.1515/cmble-2015-0010

We previously characterized the β-actin gene promoter of Indian domesticated rohu carp (Labeo rohita) and made a reporter construct via fusion to green fluorescence protein (GFP) cDNA. In this study, the same construct was used to breed transgenic rohu fish. About 20% of the transgenic offspring showed ubiquitous expression of the reporter GFP gene. In a few of the transgenic fish, we documented massive epithelial and/or muscular expression with visible green color under normal light. The expression of GFP mRNA was higher in the muscle tissue of transgenic fish than in that of non-transgenic fish. A highly efficient nucleofection protocol was optimized to transfect proliferating spermatogonial cells of rohu using this reporter construct. The β-actin promoter also drove expressions in HEK293 (derived from human embryonic kidney cells), K562 (human leukemic cells) and SF21 (insect ovarian cells) lines. These findings imply conserved regulatory mechanisms of β-actin gene expression across eukaryotes. Furthermore, the isolated β-actin promoter with consensus regulatory elements has the potential to be used in generating transgenic carp with genes of interest and in basic biology research.