New non-viral method for gene transfer into primary cells.
The availability of genetically altered cells is an essential prerequisite for many scientific and therapeutic applications including functional genomics, drug development, and gene therapy. Unfortunately, the efficient gene transfer into primary cells is still problematic. In contrast to transfections of most cell lines, which can be successfully performed using a variety of methods, the introduction of foreign DNA into primary cells requires a careful selection of gene transfer techniques. Whereas viral strategies are time consuming and involve safety risks, non-viral methods proved to be inefficient for most primary cell types. The Nucleofector technology is a novel gene transfer technique designed for primary cells and hard-to-transfect cell lines. This non-viral gene transfer method is based on a cell type specific combination of electrical parameters and solutions. In this report, we show efficient transfer of DNA expression vectors and siRNA oligonucleotides into a variety of primary cell types from different species utilizing the Nucleofector technology, including human B-CLL cells, human CD34+ cells, human lymphocytes, rat cardiomyocytes, human, porcine, and bovine chondrocytes, and rat neurons.
Gresch
Oliver
O
amaxa GmbH, Cologne, Germany.
Engel
Felix B
FB
Nesic
Dobrila
D
Tran
Thi Thai
TT
England
Hazel M
HM
Hickman
Emma S
ES
Körner
Ida
I
Gan
Li
L
Chen
Sylvia
S
Castro-Obregon
Susana
S
Hammermann
Rainer
R
Wolf
Jürgen
J
Müller-Hartmann
Herbert
H
Nix
Michael
M
Siebenkotten
Gregor
G
Kraus
Günter
G
Lun
Klaus
K
D000818:Animals; D002318:Cardiovascular Diseases / Q000628:therapy; D002417:Cattle; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D032342:Muscle Cells / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D014162:Transfection / Q000379:methods*; D014780:Viruses / Q000235:genetics
D034741:RNA, Small Interfering