Literature DB >> 15121166

Biolistic and diolistic transfection: using the gene gun to deliver DNA and lipophilic dyes into mammalian cells.

John O'Brien1, Sarah C R Lummis.   

Abstract

Delivery of genes by firing them into mammalian cells is becoming increasingly popular for cells such as neurones, that have proved difficult to transfect by conventional means. In addition it is becoming apparent that this technique also provides a suitable method of introducing cell-specific dyes into mammalian tissues. Here we describe improved protocols for the rapid and efficient delivery of both DNA and lipophilic dyes into mammalian cells in both monolayers and in brain slices. The results show that transfection of genes, such as that for enhanced yellow fluorescent protein, using a modified microcarrier preparation technique combined with either a conventional or novel accelerator channel, yields rapid and efficient production of protein, which can be visualised in 24-48 h. Following similar delivery of lipophilic dyes neurones can be visualised in minutes. These techniques therefore provide an excellent means not only to examine the structure and function of neurones and neuronal genes, but also those of other cell types. Copyright 2003 Elsevier Inc.

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Year:  2004        PMID: 15121166     DOI: 10.1016/j.ymeth.2003.11.010

Source DB:  PubMed          Journal:  Methods        ISSN: 1046-2023            Impact factor:   3.608


  5 in total

Review 1.  Physical methods of nucleic acid transfer: general concepts and applications.

Authors:  Julien Villemejane; Lluis M Mir
Journal:  Br J Pharmacol       Date:  2009-01-21       Impact factor: 8.739

2.  Ballistic delivery of dyes for structural and functional studies of the nervous system.

Authors:  Wen-Biao Gan; Jaime Grutzendler; Rachel O Wong; Jeff W Lichtman
Journal:  Cold Spring Harb Protoc       Date:  2009-04

3.  DiOlistics: delivery of fluorescent dyes into cells.

Authors:  Nyssa Sherazee; Veronica A Alvarez
Journal:  Methods Mol Biol       Date:  2013

4.  Focal macromolecule delivery in neuronal tissue using simultaneous pressure ejection and local electroporation.

Authors:  Matthew Barker; Brian Billups; Martine Hamann
Journal:  J Neurosci Methods       Date:  2008-10-30       Impact factor: 2.390

Review 5.  Diolistics: incorporating fluorescent dyes into biological samples using a gene gun.

Authors:  John A O'Brien; Sarah C R Lummis
Journal:  Trends Biotechnol       Date:  2007-10-22       Impact factor: 19.536

  5 in total

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