Colon epithelium can be transiently transfected with liposomes, calcium phosphate precipitation and DEAE dextran in vivo.
BACKGROUND
The introduction of recombinant DNA into cells is the initial step toward the development of gene therapy. It has been shown that cationic liposomes are useful vehicles to introduce DNA into colon epithelial cells in vivo.


METHODS
In the present study we compared the efficacy of different nonviral transfection methods into the colon wall. In anesthetized rats, a double balloon catheter was advanced into the colon and a chloramphenicol acetyltransferase (CAT) reporter plasmid complexed to liposomes, mixed with DEAE dextran, or precipitated with calcium phosphate was instilled. Following 2 days CAT activity was determined in the transfected colon segments.


RESULTS
DEAE dextran and liposomes were more effective than calcium phosphate, whereas naked DNA was not taken up by the colon epithelial cells. Reporter gene expression was dose-dependent. Expressing cell types did not differ utilizing the various transfection methods as judged by X-gal staining of colon sections after transfection with a LacZ reporter plasmid.


CONCLUSIONS
These data indicate that in addition to liposomes, plasmid DNA mixed with DEAE dextran can be taken up by colon epithelial cells. This transfection techniques may prove useful in the development of gene therapy approaches for colon disease.
Liptay
S
S
Department of Pediatrics, University of Ulm, Germany.
Weidenbach
H
H
Adler
G
G
Schmid
R M
RM
D000818:Animals; D002130:Calcium Phosphates / Q000737:chemistry / Q000494:pharmacology*; D002404:Catheterization; D011232:Chemical Precipitation; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis / Q000235:genetics; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D003637:DEAE-Dextran / Q000737:chemistry / Q000494:pharmacology*; D004247:DNA / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006651:Histocytochemistry; D008081:Liposomes / Q000737:chemistry / Q000494:pharmacology*; D008722:Methods; D010957:Plasmids / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection / Q000187:drug effects / Q000235:genetics
D002130:Calcium Phosphates; D008081:Liposomes; D011993:Recombinant Fusion Proteins; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; D004247:DNA; D003637:DEAE-Dextran; C020243:calcium phosphate; D015500:Chloramphenicol O-Acetyltransferase; C485829:calcium phosphate, dibasic, anhydrous