Non-Viral in Vitro Gene Delivery: It is Now Time to Set the Bar!

Abstract

Transfection by means of non-viral gene delivery vectors is the cornerstone of modern gene delivery. Despite the resources poured into the development of ever more effective transfectants, improvement is still slow and limited. Of note, the performance of any gene delivery vector in vitro is strictly dependent on several experimental conditions specific to each laboratory. The lack of standard tests has thus largely contributed to the flood of inconsistent data underpinning the reproducibility crisis. A way researchers seek to address this issue is by gauging the effectiveness of newly synthesized gene delivery vectors with respect to benchmarks of seemingly well-known behavior. However, the performance of such reference molecules is also affected by the testing conditions. This survey points to non-standardized transfection settings and limited information on variables deemed relevant in this context as the major cause of such misalignments. This review provides a catalog of conditions optimized for the gold standard and internal reference, 25 kDa polyethyleneimine, that can be profitably replicated across studies for the sake of comparison. Overall, we wish to pave the way for the implementation of standardized protocols in order to make the evaluation of the effectiveness of transfectants as unbiased as possible.

Characterization of iRGD-Ligand Modified Arginine-Histidine-Rich Peptides for Nucleic Acid Therapeutics Delivery to αvβ3 Integrin-Expressing Cancer Cells

“…It is important to notice that a general association between cytotoxicity and transfection efficiency suggests that a degree of membrane-damage is inevitable when effective carriers mediate DNA access to the cytoplasm. Successful transfection can rely on the correct balance between gaining the required access of DNA into the cytoplasm and causing excessive cell damage [ 32 , 33 ].…”

Recent advances in polymeric biomaterials-based gene delivery for cartilage repair

“…There are growing evidences that gene delivery using synthetic polymers for cartilage repair is a promising option for cartilage repair ( Table 1 ). More details about gene delivery non-polymeric vectors can refer to related reviews [ [17] , [18] , [19] , [24] , [25] , [26] ]. In the following section, we briefly introduce the polymeric vectors involved in gene delivery for cartilage repair, with more focus on polymer-mediated.…”

Scaffold-Mediated Gene Delivery for Osteochondral Repair

“…Gene transfer via nonviral vectors (transfection) is the incorporation of the DNA plasmid (pDNA) either alone or complexed with cationic or ionizable lipids (lipoplexes), cationic polymers (polyplexes), or a combination of both (lipopolyplexes) [ 35 ] into the target cell population [ 36 ] ( Table 1 ). More recent approaches also include the use of niosomes (nioplexes) [ 37 ], dendrimers (dendriplexes) [ 38 ], and gold or carbon nanostructures [ 39 ].…”