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. 2021 May 10;14(9):2467.
doi: 10.3390/ma14092467.

N-[4-(N,N,N-Trimethylammonium)Benzyl]Chitosan Chloride as a Gene Carrier: The Influence of Polyplex Composition and Cell Type

Sergei V Raik  1 , Tatiana V Mashel  2 , Albert R Muslimov  3   4   5 , Olga S Epifanovskaya  3 , Mikhail A Trofimov  4 , Daria N Poshina  1 , Kirill V Lepik  3 , Yury A Skorik  1
Affiliations

N-[4-(N,N,N-Trimethylammonium)Benzyl]Chitosan Chloride as a Gene Carrier: The Influence of Polyplex Composition and Cell Type

Sergei V Raik et al. Materials (Basel). .

Abstract

Polyplex-based gene delivery systems are promising substitutes for viral vectors because of their high versatility and lack of disadvantages commonly encountered with viruses. In this work, we studied the DNA polyplexes with N-[4-(N,N,N-trimethylammonium)benzyl]chitosan chloride (TMAB-CS) of various compositions in different cell types. Investigations of the interaction of TMAB-CS with DNA by different physical methods revealed that the molecular weight and the degree of substitution do not dramatically influence the hydrodynamic properties of polyplexes. Highly substituted TMAB-CS samples had a high affinity for DNA. The transfection protocol was optimized in HEK293T cells and achieved the highest efficiency of 30-35%. TMAB-CS was dramatically less effective in nonadherent K562 cells (around 1% transfected cells), but it was more effective and less toxic than polyarginine.

Keywords: cell transfection; chitosan; gene delivery; polyplex.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Intercalated EtBr fluorescence quenching: (a) pDNA, (b) salmon sperm DNA. Data presented as mean ± confidence interval (p = 0.05; n = 100). Legend is applicable for both graphs.
Figure 2
Figure 2
Hydrodynamic radii and ζ-potentials of polyplexes prepared in deionized water (a) and PBS (b).
Figure 3
Figure 3
Gel retardation assay for TMAB-CS:DNA polyplexes with different mass ratios (1 = 0.2:1; 2 = 2:1; 3 = 20:1).
Figure 4
Figure 4
Fraction of GFP expressing HEK293T cells after transfection with TMAB-CS37-54 polyplexes: (a) the amount of DNA was 50 μg/106 cells; (b) the TMAB-CS:DNA mass ratio was 25:1.
See this image and copyright information in PMC

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