Ahmad M. Khalil
Background: Integrating transgene into the host genome for stable maintenance in dividing cells can have unpredictable effects on gene expression and unintended effects on neighboring genes. Ethical concerns are particularly serious in the case of clinical applications using germ cells of heritable genome editing in humans, i.e., modifications introduced in sperm, eggs, or embryos to “create” genetically engineered children. One major challenge that faces in vivo gene editing (GE) technology is the selection of a suitable delivery route to transfer engineered DNA, RNA, or proteins into target cells. This problem originates from the large size of the key components and the low capacity of the delivery vector. In general, cell delivery methods available today can be broadly classified into three categories: chemical, biological and physical.
Methods and findings: The PubMed search was used with each keyword of cell delivery strategies regarding the description of various delivery methodologies and factors that influence transfection efficiency. Due to the exponential growth of papers that appear in the literature in the field and space limitations, the search was limited to data published in the year 2015 up to March 2022. Although the choice of the appropriate cell delivery route may look trivial, it is actually a crucial factor that is often overlooked. Selecting the origin and cell type, as well as proper experimental protocol, is of at most importance to maximize outcomes.
Conclusion: The reader should keep in mind that a side by side comparison of the different methods may be necessary to find the most suitable methodology for a specific goal. No single approach can be applied to all cells and all experiments. The transfer of macromolecules through different cellular routes is much more challenging and technically complex. Hurdles remain to overcome before opening a prospective future in pharmaceutics and gene therapy.
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