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Effect of degree of deacetylation of chitosan on gene transfection efficiency in vitro

机译：壳聚糖脱乙酰化程度对体外基因转染效率的影响

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Chitosan, a natural, biodegradable polysaccharide found in crustacean shells, has found use in many biological and medical applications. Chitosan-DNA nanospheres have shown promising results in DNA vaccination in vivo (1). Modification to the nanospheres after synthesis, in order to improve gene transfection efficiency, has also been reported (2). Despite the potential of chitosan as a gene carrier, particularly in oral delivery, it is inefficient in many other tissues. In an attempt to understand the rate-limiting steps in transfection by these DNA nanoparticles, intracellular trafficking studies conducted in our lab have led to the observation that even after 24 hours post-internalization, some plasmid was still not released from the chitosan nanospheres intracellularly (3). Hypothesizing that excessive stability of the DNA nanospheres may be a detriment to efficient transfection, we investigated parameters that might lead to a less stable formulation. Decreasing the degree of deacetylation (DA) may facilitate DNA release from the nanospheres by increasing the chitosan degradation rate and reducing the number of free amines with which the phosphates in DNA can interact. This study looks at the effect of degree of deacetylation of chitosan on transfection efficiency in vitro.
机译：壳聚糖，甲壳动物贝壳中发现的天然，可生物降解多糖，发现许多生物和医学应用。壳聚糖-DNA纳米球已经在体内（1）中所示的DNA疫苗接种有前途的结果。修改合成后的纳米球，以提高基因转染效率，也有报道（2）。尽管壳聚糖作为基因载体，特别是在口服递送的潜力，它在许多其它组织低效的。在试图了解这些DNA纳米粒子转染的限速步骤，在我们的实验室进行细胞内运输的研究已经导致了观察，即使后24小时后内，一些质粒仍然没有从壳聚糖纳米球释放细胞内（ 3）。假设到的DNA纳米球的过分稳定可以是无损于有效的转染中，我们调查了可能导致不太稳定的制剂的参数。降低脱乙酰（DA）的程度可以通过增加脱乙酰壳多糖的降解率，并减少与在DNA中的磷酸盐可相互作用的游离胺的数量有利于从纳米球DNA释放。这项研究着眼于壳聚糖脱乙酰度的体外转染效率的影响。

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