Study on influencing factors in efficiency of introducing gene into human keratinocyte (KC) / 中华烧伤杂志
Chinese Journal of Burns
;
(6): 122-125, 2009.
Artigo
em Chinês
| WPRIM
| ID: wpr-257432
ABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of plasmids in different size and gene transfection protocol on efficiency of introducing gene into human KC.</p><p><b>METHODS</b>Four plasmids in different size, inclu-ding pSUPER-enhanced green fluorescent protein (EGFP), pEGFP-N2, pHSER-green fluorescent protein (GFP) and ploxP-EGFP, were transfected into immortal human KC line (HaCaT) and human embryo kid-ney cell line (293FT) separately following transfection protocols of liposome (LTP), cation polymerizer (CPTP), electroporation combined with nucleus transfection agent (ETP) and lentivirus. 293FT was used as control. GFP expression was observed under inverted fluorescence microscope. The transfection efficiency (TE) was calculated.</p><p><b>RESULTS</b>(1) The four plasmids could be introduced into HaCaT (TE, 1.0%-3.3%) and 293FT (TE, 80.0%-84.7% ) following LTP. (2) The four plasmids could also be introduced into HaCaT (TE, 1.0%-3.7% ) and 293FT (TE, 81.3%-86.7% ) following CPTP. (3) Two shorter plasmids (pSUPER-EGFP and pEGFP-N2) could be introduced into HaCaT by ETP with higher TE than the othr two longer plasmids (pHSER-GFP and ploxP-EGFP), which were 22.3% and 19.0% vs. 4.0% and 3.3%, respectively. (4) pHSER-GFP packaged by lentivirus could be introduced into HaCaT with the TE reaching 97.0%, which surpassed the above three protocols.</p><p><b>CONCLUSIONS</b>It is difficult to introduce exogenous gene into human KC by LTP or CPTP; TE of lentivirus transfection protocol apparently surpasses</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Transfecção
/
Terapia Genética
/
Queratinócitos
/
Linhagem Celular
/
Vetores Genéticos
/
Lipossomos
/
Metabolismo
/
Métodos
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Burns
Ano de publicação:
2009
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS