TY  - JOUR
AU  - Nundkumar, Nirasha
AU  - Singh, Sooboo
AU  - Singh, Moganavelli
PY  - 2023
DA  - 2023/09/21
TI  - Arginine and Histidine-modified Layered Double Hydroxides Facilitate Transgene Expression in Cancer Cells in Vitro
JO  - OBM Genetics
SP  - 193
VL  - 07
IS  - 03
AB  - Layered double hydroxides (LDHs) have interesting properties and structures that enable them to carry nucleic acids, such as deoxyribonucleic acid (DNA). This study synthesized LDHs using the co-precipitation method and functionalized with the amino acids arginine (Arg) and histidine (His) to promote proton-sponge activity for enhanced transgene expression. The LDHs were characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM), and nanoparticle tracking analysis (NTA). The interaction of the LDHs with the reporter gene plasmid DNA (pCMV-Luc DNA) was determined using agarose gel electrophoresis. Cytotoxicity and transgene expression was assessed using the 3-(4, 5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) and luciferase reporter gene assay in the human embryonic kidney (HEK293), colorectal carcinoma (Caco-2) and hepatocellular carcinoma (HepG2) cells. The DNA: LDH complexes were relatively non-cytotoxic to all cells, and the highest transgene expression was achieved in the HEK293 cells exhibiting the most significant degree of transfection, followed by the Caco-2 cells. The His-LDH complexes displayed more than a two-fold increase in transfection than the Arg-LDHs, especially in the HEK293 cells at the optimal binding ratio. The non-functionalized LDHs demonstrated high transfection, which exceeded that of the His-LDH and Arg-LDH by 20% and 30%, respectively, in the Caco-2 cells. Little difference was noted in the HepG2 cells, which presented with the lowest transfection. These LDHs have demonstrated the potential to bind, protect, and efficiently deliver pDNA in vitro.
SN  - 2577-5790
UR  - https://doi.org/10.21926/obm.genet.2303193
DO  - 10.21926/obm.genet.2303193
ID  - Nundkumar2023
ER  -