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There is no single or universal transfection strategy that is apt for all cell lines and experimental aims. Apart from the experimental budget and the availability of required facilities, the choice for an optimal transfection approach or strategy depends on factors including the type and origin of cells and the form of nucleic acids to be transfected. It is also essential to consider factors that may affect transfection efficiency and cytotoxicity to the host cells and how these parameters may be assessed accurately and conveniently.
Likewise, the inclusion of appropriate controls in a transfection experiment is equally important to allow a fair and unbiased assessment of the experimental outcome. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests The authors declare that they have no competing interests. Data Availability The following information was supplied regarding data availability:. Published online Apr Author information Article notes Copyright and License information Disclaimer.
Corresponding author. Wan Yong Ho: ym. Received Jul 22; Accepted Mar 5. This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. Abstract Transfection is a modern and powerful method used to insert foreign nucleic acids into eukaryotic cells.
Open in a separate window. Figure 1. Transfection approaches Transfection methods can be generally divided into viral, non-viral, or a combination of both hybrid Fig. Figure 2. Different transfection protocols that can be divided into viral-based, non-viral based or combination of both hybrid.
Viral transfection Viral-based transfection, or more specifically known as transduction, involves using a viral vector to carry a specific nucleic acid sequence into a host cell.
Figure 3. Summary of the commonly used chemical transfection reagents. Hybrid of viral and non-viral transfection methods The combination of viral and non-viral transfection method is known as a hybrid approach, and the use of such a combined method was reported to produce higher transfection efficiency compared to other transfection methods such as using polyplexes alone Pinnapireddy et al.
Type of transfected nucleic acids Deoxyribonucleic acids In transfection, DNAs are normally transported into a host cell via a viral or non-viral vector such as plasmid Balak et al.
Table 1 List of commonly used oligonucleotides in the small RNA transient transfection work. Combination of different transfected nucleic acids or co-transfection Co-transfection is a process in which more than one type of nucleic acid is being introduced into the eukaryotic cell Fig. Figure 4. Applications of co-transfection in biotechnology and life sciences research.
Assessing transfection efficiency Assessing the efficiency of transfection is vital, especially in functional studies which require high transfection efficiencies to warrant post-transcriptional regulation of specific downstream targets Weilin et al. Table 2 Comparison of the different methods for the assessment of transfection efficiency.
Different approaches for assessing transfection efficiency. Factors influencing transfection efficiency The efficiency of chemical transfection depends greatly on a few factors such as type of reagents used, the origin and nature of target cells, and an optimum DNA to reagent ratio chosen Table S1 Gharaati-Far et al. Factors influencing the efficiency of viral transfection Viral vectors such as lentiviruses are useful in gene therapy due to their ability to carry large-sized nucleic acid and deliver their targets to both non-dividing and dividing cells Karda et al.
Type of chemical transfection reagents Primary and stem cells Primary mammalian cells are generally less susceptible to transfection than other cell types due to its finite lifespan and limited expansion capacity Oh et al. Human vs animal cells The origin of cell lines, such as human vs. Cell lines derived from the same species The performance of different transfection reagents could also vary in different cell lines derived from the same species.
Adherent vs. The ratio of nucleic acids to transfection reagents The ratio of nucleic acids to transfection reagents also plays a role in influencing transfection efficiency Arnold et al. Other factors Serum-reduced or serum-free media are normally recommended in transfection involving cationic transfection reagents such as Lipofectamine Wallenstein et al. Conclusion There is no single or universal transfection strategy that is apt for all cell lines and experimental aims.
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