An efficient gene transfer system for hematopoietic cell line using transient and stable vectors.
The hematopoietic system represents an interesting model for gene transfer protocols. Here, we have evaluated the efficiency of a gene transfer system using the polycationic compound SuperFect (Qiagen) and the K562 hematopoietic cell line. Transient and stable vectors carrying the enhanced green fluorescent protein (EGFP) reporter gene were employed. The stable vector was constructed based on Epstein-Barr virus sequences such as EBV oriP (origin of replication) and EBNA (EBV nuclear antigen)-1, both for DNA replication. The transfection efficiency of the viable cells was estimated by flow cytometry at approximately 98% for transient and stable vectors. Transiently transfected cells presented optimal EGFP expression until day 2 when fluorescence started to decrease. In contrast, stable transfectants continuously expressed the marker gene product for 10 weeks in the presence of G418. Our results represent an efficient gene transfer method for K562 hematopoietic cells and may be used as an alternative approach for further gene transfer studies involving hematopoietic cells.
Teixeira
L A
LA
Departamento de Genética, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500 C.P. 15053, 91501-970, RS, Porto Alegre, Brazil. teixeira@if.ufrgs.br
Fricke
C H
CH
Bonorino
C B
CB
Bogo
M R
MR
Nardi
N B
NB
D002460:Cell Line; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D018741:Replication Origin / Q000235:genetics
D019309:Epstein-Barr Virus Nuclear Antigens; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; C417029:EBV-encoded nuclear antigen 1