Preparation and characterization of DET-CPD-12 polyplexes.
Agarose
gel electrophoresis assay of DET-CPD-12 polyplexes with Cas9 plasmid
(a) and Cas9 mRNA (d) at different N/P ratios. Particle size and ζ
potential of DET-CPD-12/Cas9 plasmid complexes (b), DET-CPD-12/Cas9
mRNA complexes (e), and DET-CPD-12/Cas9 protein complexes (h). The
inset is the TEM image of the nanoparticles, and all of the scale
bars represent 200 nm. Percentage of free genome-editing biomacromolecules
after the formation of DET-CPD-12/Cas9 DNA complexes (c), DET-CPD-12/Cas9
mRNA complexes (f), and DET-CPD-12/Cas9 protein complexes (i). The
release of Cas9 DNA, mRNA, and protein from their complexes was also
evaluated in the presence or absence of 10 mM GSH. All data represent
mean ± S.D. (n = 3). (g) SDS-page assay of DET-CPD-12/Cas9
protein complexes at different weight ratios.