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

Sergei V Raik,Albert R Muslimov,Yury A Skorik,Mikhail A Trofimov,Kirill V Lepik,Olga S Epifanovskaya,Daria N Poshina,Tatiana V Mashel

doi:10.3390/ma14092467

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.

Highlights

Published: 10 May 2021Gene therapy is a therapeutic approach that involves the introduction of nucleic acids into a patient’s cells to correct disease
The modification of CS amino groups with a TMAB substituent by reductive alkylation is highly efficient at relatively low degrees of substitution (DS)
A two-fold amount of FTMA led to a DS higher than 50%

Summary

Introduction

Gene therapy is a therapeutic approach that involves the introduction of nucleic acids into a patient’s cells to correct disease It is an extensively evolving field of medicine that holds significant promise for the treatment of conditions presently considered incurable by conventional approaches, and it is currently being tested in more than 3000 clinical trials worldwide [1,2]. Despite this potential, the introduction of novel gene therapies into clinical practice remains hampered by a lack of efficient and safe gene delivery methods [3,4]. ZolgensmaTM is another that is indicated for the treatment of spinal muscular atrophy [6]

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