Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralize

• Autor(es): Groll, Andrea Von ; Levin, Yan ; Barbosa, Marcia Cristina Bernardes ; Ravazzolo, Ana Paula
• Data: 2013
• Origem: Oasisbr
• Tipo de Documento: Artigo científico
• Idioma: Inglês

Descrição

Submitted by cristiane soares ([e-mail]) on 2012-09-17T00:38:57Z No. of bitstreams: 1 Linear DNA Low Efficiency Transfection by Liposome Can Be Improved by the Use of Cationic Lipid as Charge Neutralize.pdf: 371951 bytes, checksum: 0a03e7add6e09a93bb9d608bdac0925a (MD5)

Approved for entry into archive by Gabriela Silva da Rosa([e-mail]) on 2013-01-28T22:53:08Z (GMT) No. of bitstreams: 1 Linear DNA Low Efficiency Transfection by Liposome Can Be Improved by the Use of Cationic Lipid as Charge Neutralize.pdf: 371951 bytes, checksum: 0a03e7add6e09a93bb9d608bdac0925a (MD5)

Made available in DSpace on 2013-01-28T22:53:08Z (GMT). No. of bitstreams: 1 Linear DNA Low Efficiency Transfection by Liposome Can Be Improved by the Use of Cationic Lipid as Charge Neutralize.pdf: 371951 bytes, checksum: 0a03e7add6e09a93bb9d608bdac0925a (MD5) Previous issue date: 2006

A plasmid expressing the â-galactosidase enzyme was used to transfect Vero cells in order to evaluate the efficiency of a liposome-mediated transfection by circular and linear DNA. The results obtained showed a low rate of transfection by linear DNA:liposome complexes. To explore whether the structure of the complexes was interfering with the transfection, atomic force microscopy (AFM) was used. It has confirmed the difference between the linear and circular condensates: whereas the circular DNA:liposome complexes presented compact spherical or cylindrical structures of about 100-800 nm, the linear DNA showed pearl necklace-like structures,with pearls varying from 250 to 400 nm. On the basis of the theory proposed by Kuhn et al. (1999), low concentrations of cationic amphihile were used to neutralize or reverse the DNA charge in order to improve the transfection efficiency of the linear DNA. Using this method,we were able to obtain the expression of the transgene without an associated toxicity observed with the linear DNA liposome delivery.