Development of human gene reporter cell lines using rAAV mediated homologous recombination

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Author: Fernandez, SL
Author: Hurlin, PJ
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Methods Article

Development of human gene reporter cell lines using rAAV mediated homologous recombination

Sandra L. Fernandez¹, David W. Russell² and Peter J. Hurlin³^*

¹ Shriners Hospitals for Children. Department of Cell Developmental Biology, Oregon Health & Science Portland. OR 97201. USA.
² Department of Pediatrics, Division of Genetics and Developmental Medicine, University of Washington Seattle. WA 98195-7720. USA.
³ Department of Cell Developmental Biology, Oregon Health & Science Portland. OR 97201. USA. Phone: (503) 221 3438 Fax: (503) 221 3451 Email: pjh@shcc.org.

* To whom correspondence should be addressed: Peter J. Hurlin, Department of Cell Developmental Biology, Oregon Health & Science Portland. OR 97201. USA. Phone: (503) 221 3438. Fax: (503) 221 3451. Email: pjh@shcc.org

Biol. Proced. Online 2007;9:84-90. doi:10.1251/bpo136
Submitted: September 10, 2007; Accepted: November 19, 2007; Published: December 24, 2007.

Indexing terms: Dependovirus; Proto-Oncogene proteins c-myc.

Abbreviations: EGFP, enhanced green fluorescent protein; HFF, human foreskin fibroblast; rAAV, recombinant adeno-associated virus.

Figure 4 Enlarged

Fig. 4:

Comparison of c-Myc and EGFP-Luciferase reporter gene expression and activity kinetics in response to cell cycle entry. The MR1 cells were tracked through CCE for their ability to induced c-Myc and EGFP-Luciferase transcripts by RT-PCR (A) and protein by Western blot (B). Max protein expression is used as a loading control. (C) Luciferase activity of the MR1 population over CCE. Data shown is the average of triplicate samples +/- SEM. (D) FACS Calibur data is shown where the EGFP-Luciferase positive percent of the parent population was determined using BD CellQuest Pro v. 5.2. Time course changes of the percent EGFP-Luciferase expressing populations were compared under the same gates.

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