Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma

Ömer Aktürk; Beste Çağdaş Tunalı

doi:10.29137/umagd.943584

EN TR

A Study On The siRNA Transfection Reagent Potential of Levan-Capped Gold Nanoparticles Synthesized by Laser Ablation

Abstract

Here, gold nanoparticle (AuNP)-based small interfering RNA (siRNA) vector was developed. AuNPs synthesized by pulsed laser ablation in liquid (PLAL) method in a medium containing levan polysaccharide (Levan-AuNP) were characterized, then utilized in siRNA transfection studies on MCF-7 breast cancer cells. The interaction between non-targeted siRNA sequence and Levan-AuNP-siRNA was found to be 48% via A260 absorbance measurements. Optimal conditions for siRNA transfection in MCF-7 cells were determined by preparing different concentrations of Levan-AuNP and 50 nM siGLO Green transfection indicator. In transfection studies, initially triple fluorescent staining protocols (siGLO Green, propidium iodide (PI), and Hoechst 33342) were conducted and no cytotoxicity was observed. In 1 day-incubation, the transfection was realized especially in the highest concentration Levan-AuNP group while it was unsuccessful in siGLO Green group alone. Transfection was observed in all groups after 2 and 4 days of incubation. In the secondary transfection studies without PI and Hoechst, the fluorescence intensity of siGLO Green increased significantly, and stronger and punctuate fluorescence emission was seen in Levan-AuNP-siGLO Green complexes. Regarding the mean fluorescence intensity measurements, a statistical significance was confirmed in all Levan-AuNP groups. As a whole, the developed Levan-AuNP could be proposed as alternatives to the existing transfection agents.

Keywords

Levan, gold nanoparticle, laser, siRNA, transfection

Project Number

2017/017

Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma

Abstract

Bu çalışmada, altın nanoparçacık (AuNP) temelli kısa interferanslı RNA (siRNA) transfeksiyon vektörü tasarlanmıştır. Sıvı içinde atımlı lazer ablasyon yöntemiyle (PLAL) levan polisakkariti içeren ortamda sentezlenen AuNP’ler (Levan-AuNP), karakterizasyon testlerinin ardından MCF-7 meme kanseri hücrelerinde siRNA transfeksiyon çalışmalarında kullanılmıştır. Hedefsiz siRNA sekansı ile gerçekleşen %48’lik Levan-AuNP-siRNA etkileşimi A260 absorbans ölçümleriyle bulunmuştur. Farklı derişimlerde Levan-AuNP ve 50 nM siGLO Green transfeksiyon indikatörü hazırlanarak MCF-7 hücrelerindeki optimum siRNA transfeksiyon koşulları belirlenmiştir. Transfeksiyon çalışmalarında öncelikle üçlü floresan boyama (siGLO Green, propidyum iyodür (PI) ve Hoechst 33342) yapılmış ve sitotoksisite görülmemiştir. Özellikle en yüksek derişimdeki Levan-AuNP grubunda ilk günde transfeksiyon gerçekleşirken, tek başına siGLO Green grubunda transfeksiyon gerçekleşmemiştir. 2 ve 4 günlük inkübasyonda ise tüm gruplarda transfeksiyon gözlemlenmiştir. PI ve Hoechst kullanılmadan yapılan ikinci transfeksiyon çalışmalarında ise siGLO Green floresan şiddetinin belirgin şekilde arttığı ve özellikle Levan-AuNP-siGLO Green komplekslerinde daha kuvvetli ve noktalı bir floresan ışıma görülmüştür. Ortalama floresan şiddetleri ölçümlerine göre, tüm Levan-AuNP gruplarında istatistiksel açıdan anlamlı ölçüde artış bulunmuştur. Sonuç olarak, bu çalışmada geliştirilen Levan-AuNP’nin, mevcut transfeksiyon ajanlarına alternatif olabileceği gösterilmiştir.

Keywords

Levan, altın nanoparçacık, lazer, siRNA, transfeksiyon

Supporting Institution

Kırıkkale Üniversitesi Bilimsel Araştırmalar Proje Koordinasyon Birimi

Project Number

2017/017

Thanks

Bu proje Kırıkkale Üniversitesi Bilimsel Araştırmalar Proje Koordinasyon Birimi tarafından desteklenmiştir (No: 2017/017).

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Details

Primary Language

Turkish

Subjects

Engineering

Journal Section

Research Article

Authors

Ömer Aktürk
0000-0002-0130-8803
Türkiye

Beste Çağdaş Tunalı ^*
0000-0003-2534-382X
Türkiye

Publication Date

January 31, 2022

Submission Date

May 27, 2021

Acceptance Date

September 2, 2021

Published in Issue

Year 2022 Volume: 14 Number: 1

DOI

https://doi.org/10.29137/umagd.943584

IZ

https://izlik.org/JA56GZ86NP

APA

Aktürk, Ö., & Çağdaş Tunalı, B. (2022). Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma. International Journal of Engineering Research and Development, 14(1), 282-298. https://doi.org/10.29137/umagd.943584

AMA

1.Aktürk Ö, Çağdaş Tunalı B. Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma. IJERAD. 2022;14(1):282-298. doi:10.29137/umagd.943584

Chicago

Aktürk, Ömer, and Beste Çağdaş Tunalı. 2022. “Lazer Ablasyonu Ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların SiRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma”. International Journal of Engineering Research and Development 14 (1): 282-98. https://doi.org/10.29137/umagd.943584.

EndNote

Aktürk Ö, Çağdaş Tunalı B (January 1, 2022) Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma. International Journal of Engineering Research and Development 14 1 282–298.

IEEE

[1]Ö. Aktürk and B. Çağdaş Tunalı, “Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma”, IJERAD, vol. 14, no. 1, pp. 282–298, Jan. 2022, doi: 10.29137/umagd.943584.

ISNAD

Aktürk, Ömer - Çağdaş Tunalı, Beste. “Lazer Ablasyonu Ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların SiRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma”. International Journal of Engineering Research and Development 14/1 (January 1, 2022): 282-298. https://doi.org/10.29137/umagd.943584.

JAMA

1.Aktürk Ö, Çağdaş Tunalı B. Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma. IJERAD. 2022;14:282–298.

MLA

Aktürk, Ömer, and Beste Çağdaş Tunalı. “Lazer Ablasyonu Ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların SiRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma”. International Journal of Engineering Research and Development, vol. 14, no. 1, Jan. 2022, pp. 282-98, doi:10.29137/umagd.943584.

Vancouver

1.Ömer Aktürk, Beste Çağdaş Tunalı. Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma. IJERAD. 2022 Jan. 1;14(1):282-98. doi:10.29137/umagd.943584

Kırıkkale University, Faculty of Engineering and Natural Science, 71450 Yahşihan / Kırıkkale, Türkiye.

ijerad@kku.edu.tr

A Study On The siRNA Transfection Reagent Potential of Levan-Capped Gold Nanoparticles Synthesized by Laser Ablation

Abstract

Keywords

Project Number

Lazer Ablasyonu ile Sentezlenmiş Levan Kaplı Altın Nanoparçacıkların siRNA Transfeksiyon Ajanı Olma Potansiyeli Üstüne Çalışma

Abstract

Keywords

Supporting Institution

Project Number

Thanks

References

Details

Primary Language

Subjects

Journal Section

Authors

Publication Date

Submission Date

Acceptance Date

Published in Issue

DOI

IZ

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