Recombinant Adeno-Associated Viral Vector Transduction of Human Prostate Cancer Cell Lines

Muhammet Burak Batir

Makale detayları

Recombinant Adeno-Associated Viral Vector Transduction of Human Prostate Cancer Cell Lines

Yazarlar:

Muhammet Burak Batir

Yayıncı:

Manisa Celal Bayar University

Ana Başlıklar:

Çözelti Kimyası

Dergi:

Celal Bayar Üniversitesi Fen Bilimleri Dergisi Vol.19 Issue 4, pp.333-341

Belge Tipi:

Makale

Yayın Tarihi:

12/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001308170

pISSN:

1305-130X

Anahtar kelimeler:

Özet:

 Abstract: At the core of gene therapy lies the use of viral vectors, engineered viruses serving as delivery vehicles to transport restorative genes into target cells. Therefore, the effect of 7 different rAAV serotypes and their different quantites was analysis here on human prostate cancer cell lines PC-3 and DU-145, which are hard to be transfected. PC-3 and DU-145 cell lines were infected with different multiplicity of infection MOI ratios of 7 rAAV serotypes, AAV 21, 22, 23, 25, 26, and 29, which were expressing the green fluorescent protein GFP transgene driven by the CMV promoter. The transduction efficiency was analyzed by fluorescent microscopy and flow cytometry. In addition, the cell viability of the infected cells was measured by Muse Cell Analyzer at the MOI of 10.000. rAAV 22 and rAAV 26 have the most significant ability to transduce PC-3 cells. Although rAAV 22 and rAAV 26 were also the most transducing serotypes in the DU-145 cell line, the transduction rates did not exceed 20 in this cell line. On the other hand, after viral infection, no difference in cell viability was observed in PC-3 cells compared to the mock group, while a significant decrease in viability was observed in DU-145 cells. This study determined the transduction efficiency of 7 different rAAV serotypes on human cancer cell lines. While rAAV 22 and rAAV 26 serotypes achieved more than 60 transduction efficiency in PC-3 cells, the transduction efficiency could not exceed 20 in DU-145 cells. Overall, this study demonstrated that rAAV 22 and rAAV 26 could mediate the expression of a transgene with a high transduction efficiency.

 Abstract: At the core of gene therapy lies the use of viral vectors, engineered viruses serving as delivery vehicles to transport restorative genes into target cells. Therefore, the effect of 7 different rAAV serotypes and their different quantites was analysis here on human prostate cancer cell lines PC-3 and DU-145, which are hard to be transfected. PC-3 and DU-145 cell lines were infected with different multiplicity of infection MOI ratios of 7 rAAV serotypes, AAV 21, 22, 23, 25, 26, and 29, which were expressing the green fluorescent protein GFP transgene driven by the CMV promoter. The transduction efficiency was analyzed by fluorescent microscopy and flow cytometry. In addition, the cell viability of the infected cells was measured by Muse Cell Analyzer at the MOI of 10.000. rAAV 22 and rAAV 26 have the most significant ability to transduce PC-3 cells. Although rAAV 22 and rAAV 26 were also the most transducing serotypes in the DU-145 cell line, the transduction rates did not exceed 20 in this cell line. On the other hand, after viral infection, no difference in cell viability was observed in PC-3 cells compared to the mock group, while a significant decrease in viability was observed in DU-145 cells. This study determined the transduction efficiency of 7 different rAAV serotypes on human cancer cell lines. While rAAV 22 and rAAV 26 serotypes achieved more than 60 transduction efficiency in PC-3 cells, the transduction efficiency could not exceed 20 in DU-145 cells. Overall, this study demonstrated that rAAV 22 and rAAV 26 could mediate the expression of a transgene with a high transduction efficiency.

 Abstract: At the core of gene therapy lies the use of viral vectors, engineered viruses serving as delivery vehicles to transport restorative genes into target cells. Therefore, the effect of 7 different rAAV serotypes and their different quantites was analysis here on human prostate cancer cell lines PC-3 and DU-145, which are hard to be transfected. PC-3 and DU-145 cell lines were infected with different multiplicity of infection MOI ratios of 7 rAAV serotypes, AAV 21, 22, 23, 25, 26, and 29, which were expressing the green fluorescent protein GFP transgene driven by the CMV promoter. The transduction efficiency was analyzed by fluorescent microscopy and flow cytometry. In addition, the cell viability of the infected cells was measured by Muse Cell Analyzer at the MOI of 10.000. rAAV 22 and rAAV 26 have the most significant ability to transduce PC-3 cells. Although rAAV 22 and rAAV 26 were also the most transducing serotypes in the DU-145 cell line, the transduction rates did not exceed 20 in this cell line. On the other hand, after viral infection, no difference in cell viability was observed in PC-3 cells compared to the mock group, while a significant decrease in viability was observed in DU-145 cells. This study determined the transduction efficiency of 7 different rAAV serotypes on human cancer cell lines. While rAAV 22 and rAAV 26 serotypes achieved more than 60 transduction efficiency in PC-3 cells, the transduction efficiency could not exceed 20 in DU-145 cells. Overall, this study demonstrated that rAAV 22 and rAAV 26 could mediate the expression of a transgene with a high transduction efficiency.

 Abstract: At the core of gene therapy lies the use of viral vectors, engineered viruses serving as delivery vehicles to transport restorative genes into target cells. Therefore, the effect of 7 different rAAV serotypes and their different quantites was analysis here on human prostate cancer cell lines PC-3 and DU-145, which are hard to be transfected. PC-3 and DU-145 cell lines were infected with different multiplicity of infection MOI ratios of 7 rAAV serotypes, AAV 21, 22, 23, 25, 26, and 29, which were expressing the green fluorescent protein GFP transgene driven by the CMV promoter. The transduction efficiency was analyzed by fluorescent microscopy and flow cytometry. In addition, the cell viability of the infected cells was measured by Muse Cell Analyzer at the MOI of 10.000. rAAV 22 and rAAV 26 have the most significant ability to transduce PC-3 cells. Although rAAV 22 and rAAV 26 were also the most transducing serotypes in the DU-145 cell line, the transduction rates did not exceed 20 in this cell line. On the other hand, after viral infection, no difference in cell viability was observed in PC-3 cells compared to the mock group, while a significant decrease in viability was observed in DU-145 cells. This study determined the transduction efficiency of 7 different rAAV serotypes on human cancer cell lines. While rAAV 22 and rAAV 26 serotypes achieved more than 60 transduction efficiency in PC-3 cells, the transduction efficiency could not exceed 20 in DU-145 cells. Overall, this study demonstrated that rAAV 22 and rAAV 26 could mediate the expression of a transgene with a high transduction efficiency.

Makale detayları

Recombinant Adeno-Associated Viral Vector Transduction of Human Prostate Cancer Cell Lines

Yazarlar:

Muhammet Burak Batir

Yayıncı:

Manisa Celal Bayar University

Ana Başlıklar:

Çözelti Kimyası

Dergi:

Celal Bayar Üniversitesi Fen Bilimleri Dergisi Vol.19 Issue 4, pp.333-341

Belge Tipi:

Makale

Yayın Tarihi:

12/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001308170

pISSN:

1305-130X

Cancer

Gene therapy

rAAV

Recombinant adeno-associated virus

Transduction

Sayı

Makale detayları

Recombinant Adeno-Associated Viral Vector Transduction of Human Prostate Cancer Cell Lines

Yazarlar: Muhammet Burak Batir

Yayıncı: Manisa Celal Bayar University

Ana Başlıklar: Çözelti Kimyası

Dergi: Celal Bayar Üniversitesi Fen Bilimleri Dergisi Vol.19 Issue 4, pp.333-341

Belge Tipi: Makale

Yayın Tarihi: 12/2023

Belge Dili: İngilizce

Document ID (Org No.): 2001308170

pISSN: 1305-130X

Cancer

Gene therapy

rAAV

Recombinant adeno-associated virus

Transduction

Sayı

Yazarlar:

Muhammet Burak Batir

Yayıncı:

Manisa Celal Bayar University

Ana Başlıklar:

Çözelti Kimyası

Dergi:

Celal Bayar Üniversitesi Fen Bilimleri Dergisi Vol.19 Issue 4, pp.333-341

Belge Tipi:

Makale

Yayın Tarihi:

12/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001308170

pISSN:

1305-130X