Ultrasound-mediated gene transfer into neuronal cells.

نویسندگان

  • Andy J Fischer
  • Jennifer J Stanke
  • Ghezal Omar
  • Candice C Askwith
  • Richard W Burry
چکیده

A new field of gene transfer is emerging as a simple, effective means to drive the expression foreign genes in cells: ultrasound-mediated gene transfer or sonoporation. We report here that sonoporation is an effective means of gene transfer for cultured neurons, a cell type that has been difficult to transfect. Neuronal cell types that are effectively sonoporated include chick retinal neurons, chick dorsal forebrain, chick optic tectum, PC12 cells, rat cerebellar neurons and mouse hippocampal neurons. Depending on the type of cell and conditions of sonoporation the transfection efficacy was as high as 20%. Sonoporation of plasmid DNA was effective for cells adherent to a substrate and for free-floating cells that were freshly dissociated. In the free-floating preparations, between 60 and 95% of the cells that were transfected were neuronal, as much as 90% higher than that observed for other methods of gene transfer including adenovirus and lipid-based transfection methods. We conclude that sonoporation is a simple, effective and inexpensive means by which to preferentially transfect DNA into neuronal cells.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Induction of Human Embryonic Stem Cells into neuronal differentiation by increasing cyclic Adenosine Mono Phosphate

Introduction: To evaluate the cAMP -mediated IBMX (3-IsoButyle -1-Methyl Xanthin) and db-cAMP (dibutyryl cAMP) effects on differentiation of human Embryonic Stem Cells (hESCs) into nerve cells were the objectives of this study. Methods: We have used Royan H1 hESC- derived embryoid bodies with four treatment groups: six days treatment with IBMX (5×10 -4M) and db-cAMP (10 -9M) (referred to as...

متن کامل

Optimization of Agrobacterium-mediated transformation in oyster mushroom (Pleurotus ostreatus) by vector containing human pro-insulin gene

Transferring foreign genes into mushroom mediated by Agrobacterium tumefaciens is a standard technique in genetic engineering. Recombinant human insulin has been greatly used in the treatment of type I diabetes. The production of edible mushroom derived insulin should facilitate oral delivery. In this study we used the Agrobacterium tumefaciens mediated transformation method for the transfer an...

متن کامل

The Demonstration of Polyethylenimine Mediated Gene Transfer into the Rodent Hypothalamus Results in Persistent Over-Expression and Phenotypic Change

Polyethylenimine (PEI) has been proposed as a non-viral vector, and has been successfully used to transfer reporter genes into the central nervous system (CNS), kidneys, and lungs of adult mice. Neuropeptide Y (NPY) is a peptide expressed in the hypothalamus and is important in the regulation of body weight. Using PEI combined with stereotactic microinjection, we have successfully transferred c...

متن کامل

Studies on neutral, cationic and biotinylated cationic microbubbles in enhancing ultrasound-mediated gene delivery in vitro and in vivo.

Ultrasound-mediated gene transfer is emerging as a practical means of facilitating targeted gene expression and is significantly enhanced in the presence of exogenously added microbubbles. This study explores the influence of microbubble surface modifications on their interaction with plasmid DNA and target cells, and the functional consequences of those interactions in terms of ultrasound-medi...

متن کامل

Avian Adeno-Associated Virus Vector Efficiently Transduces Neurons in the Embryonic and Post-Embryonic Chicken Brain

The domestic chicken is an attractive model system to explore the development and function of brain circuits. Electroporation-mediated and retrovirus (including lentivirus) vector-mediated gene transfer techniques have been widely used to introduce genetic material into chicken cells. However, it is still challenging to efficiently transduce chicken postmitotic neurons without harming the cells...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Journal of biotechnology

دوره 122 4  شماره 

صفحات  -

تاریخ انتشار 2006