An efficient method for stable transfection of mouse myogenic C2C12 cell line using a nonviral transfection approach

Mehmet Deniz AKYÜZ; Burcu Balci HAYTA; Pervin R. DİNÇER

Makale detayları

An efficient method for stable transfection of mouse myogenic C2C12 cell line using a nonviral transfection approach

Yazarlar:

Mehmet Deniz AKYÜZ | Burcu Balci HAYTA | Pervin R. DİNÇER

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2011 Vol.41 Issue 5, pp.821-825

Belge Tipi:

Makale

Yayın Tarihi:

4/2011

Belge Dili:

Türkçe

Document ID (Org No.):

2000754564

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144

Anahtar kelimeler:

Özet:

 

To describe in detail the use of the T-RExTM System (a tetracycline-regulated mammalian expression system; Invitrogen) for rapid, easy, and effective generation of stable C2C12 cell clones. Tetracycline-inducible stable cell lines are useful tools to study the function of various genes in different types of mammalian cells. However, the generation of stable cell lines is a time-consuming, technically difficult, and expensive process.   Materials and methods: Generation of a stable C2C12 cell line was performed by stable transfection of the cells with the pcDNA6/TR plasmid vector, which contains the tetracycline repressor and the blasticidin drug resistance genes. The establishment of the stable cell line and the efficiency of the tetracycline-inducible gene expression system were shown with the transient transfection of reporter plasmid pTO-EGFP and observation of the GFP expression by fluorescence microscopy.   Results: We established a stable T-RExTM C2C12 cell line that can be used for muscle research studies by transfecting the gene of interest with a second plasmid.  Conclusion: The T-RExTM system is useful in the development of stable cell lines for protein production or regulation of gene expression.

 

To describe in detail the use of the T-RExTM System (a tetracycline-regulated mammalian expression system; Invitrogen) for rapid, easy, and effective generation of stable C2C12 cell clones. Tetracycline-inducible stable cell lines are useful tools to study the function of various genes in different types of mammalian cells. However, the generation of stable cell lines is a time-consuming, technically difficult, and expensive process.   Materials and methods: Generation of a stable C2C12 cell line was performed by stable transfection of the cells with the pcDNA6/TR plasmid vector, which contains the tetracycline repressor and the blasticidin drug resistance genes. The establishment of the stable cell line and the efficiency of the tetracycline-inducible gene expression system were shown with the transient transfection of reporter plasmid pTO-EGFP and observation of the GFP expression by fluorescence microscopy.   Results: We established a stable T-RExTM C2C12 cell line that can be used for muscle research studies by transfecting the gene of interest with a second plasmid.  Conclusion: The T-RExTM system is useful in the development of stable cell lines for protein production or regulation of gene expression.

Makale detayları

An efficient method for stable transfection of mouse myogenic C2C12 cell line using a nonviral transfection approach

Yazarlar:

Mehmet Deniz AKYÜZ | Burcu Balci HAYTA | Pervin R. DİNÇER

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2011 Vol.41 Issue 5, pp.821-825

Belge Tipi:

Makale

Yayın Tarihi:

4/2011

Belge Dili:

Türkçe

Document ID (Org No.):

2000754564

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144

Key words: C2C12 cell line

stable transfection

inducible expression

tetracycline resistance gene

Sayı

Makale detayları

An efficient method for stable transfection of mouse myogenic C2C12 cell line using a nonviral transfection approach

Yazarlar: Mehmet Deniz AKYÜZ | Burcu Balci HAYTA | Pervin R. DİNÇER

Yayıncı: TUBITAK

Dergi: Turkish Journal of Medical Sciences 2011 Vol.41 Issue 5, pp.821-825

Belge Tipi: Makale

Yayın Tarihi: 4/2011

Belge Dili: Türkçe

Document ID (Org No.): 2000754564

Sıklık: İki Aylık

eISSN: 1303-6165

pISSN: 1300-0144

Key words: C2C12 cell line

stable transfection

inducible expression

tetracycline resistance gene

Sayı

Yazarlar:

Mehmet Deniz AKYÜZ | Burcu Balci HAYTA | Pervin R. DİNÇER

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2011 Vol.41 Issue 5, pp.821-825

Belge Tipi:

Makale

Yayın Tarihi:

4/2011

Belge Dili:

Türkçe

Document ID (Org No.):

2000754564

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144