Novel method of single antibiotic selection of cells containing multiple stage-specific promoter-fluorophore plasmids.
This study demonstrates a novel method by which multiple separate plasmids can be stably integrated into the genome using single antibiotic resistance for selection. This method was used to integrate three different cardiac-specific promoters driving different fluorophores into murine embryonic stem cells, allowing sequential visualization of various stages of cardiac differentiation. This method is broadly applicable to the study of cell lineage of different stem/progenitor cells.
Guddati
Achuta K
AK
Department of Internal Medicine, Harlem Hospital Center at Columbia University, New York, New York 10037, USA. ag2876@columbia.edu
Kessler
John A
JA
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism*; D005821:Genetic Techniques; D006321:Heart / Q000254:growth & development; D006801:Humans; D007211:Indoles / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008858:Microscopy, Phase-Contrast; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D009355:Neomycin / Q000494:pharmacology*; D009928:Organ Specificity / Q000187:drug effects; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D055503:Protein Multimerization / Q000187:drug effects; D018168:Receptors, Retinoic Acid / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism
D005456:Fluorescent Dyes; D007211:Indoles; D018168:Receptors, Retinoic Acid; D011987:Receptors, Steroid; C055469:ecdysone receptor; C007293:DAPI; D009355:Neomycin