Section 2 Introduction to Particle Delivery
Helios Gene Gun System | 3
Section 2
Introduction to Particle Delivery
Particle Delivery Technology
Particle bombardment is a physical method of cell transformation in which high-density subcellular sized
particles are accelerated to high velocity to carry DNA into cells. The technique was first described as a
method of gene transfer into plants (Klein et al
.
1987, 1988; McCabe et al
.
1988) and subsequently shown
to be applicable to mammalian experimental systems (Zelenin et al
.
1989; Yang et al
.
1990; Williams et
al
.
1991). Because it does not depend on specific ligand receptors and/or the biochemical features of
structural components typically present on cell surfaces, particle-mediated gene transfer can be readily
applied to a variety of biological systems. Consequently, this procedure can be used to transform such
diverse targets as bacteria (Shark et al
.
1991; Smith et al
.
1992), fungi (Armaleo et al
.
1990), and intracellular
organelles (Johnston et al
.
1988; Boynton et al
.
1988). Since it is a physical method of gene delivery, particle
bombardment also overcomes physical barriers to effective gene transfer, such as the stratum corneum of
the epidermis and the cell walls of plants. Particle bombardment is a convenient method for transforming
intact cells in culture since minimal pre- or postbombardment manipulation is necessary. In addition, this
technique is much easier and faster to perform than the tedious task of microinjection. Both transient and
stable expression are possible with particle bombardment. In addition to DNA, RNA may also be transferred
to cells by particle bombardment (Qiu, et al
.
1996). Table 1 lists some of the advantages of using particle
bombardment for in vitro and in vivo
transformation.
Table 1. Advantages of particle bombardment for in vitro
and in vivo
gene transfer.
■
Easy to use, rapid, and versatile gene delivery system
■
Works with any target cell type
■
Useful for both transient and stable expression
■
Requires only small amounts of DNA
■
No carrier DNA is needed
■
Requires only small numbers of cells
■
May obtain high levels of cotransformation
■
Large DNA fragments may be transferred
■
Enables direct intracellular delivery to many cells in the target area
■
Applicable to both in vitro and in vivo
transformation
■
No extraneous genes or proteins are delivered
The Helios Gene Gun
The Helios Gene Gun is the second instrument in Bio-Rad’s particle delivery product line. In contrast to
the PDS-1000/He System, in which the overall size of the target to be transformed is limited by the size
of the chamber, and the target tissue is subjected to a vacuum during bombardment, the Helios Gene
Gun requires no vacuum and any target accessible to the barrel can be transformed. Consequently, the
Helios Gene Gun may be used in a much wider variety of gene transfer applications and provides a tool for
both in vitro and in vivo
transformations in the research lab. Essentially any type of cells that can be made
accessible to its nozzle may be transformed.
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