Customizing Kit to Install the HR DNA Repair System in Any Cell Line.

This is a custom cell-based reporter system designed to allow the customer to screen or identify agents (drugs, natural products, small molecules, synthetics, miRNAs, and genes) that affect or impact the process of HR DNA repair in virtually any cell line. The kit uses GFP as an in vivo readout for the HR pathway.  This Kit has control HeLa cells containing the DR-HRGFP reporter already integrated.  We also provide the HR Reporter DNA plasmid (DR-HRGFP) to allow customer to insert and select the reporter into his/her cell line.  In addition, we provide an I-Sce1 expression plasmid to allow the customer to transiently transfect a DNA cleavage activity and assess HR in newly made cell line.

Who Should Buy This Kit?

R&D Labs that are interested in screening for agents, drugs, genes or gene products that specifically target the entire HR DNA repair pathway in their own cell line model.  Because DNARepairKits.com is the first to market, we offer this cell line "hosting Kit" as a novel kit or service.  The system we have engineered can be hosted by any cell line capable of being transfected, even with low efficiency.  The idea is to install the DR-GFP reporter, stably in your model line and then clone out known positives, which can then be transfect with the megaendonuclease restriction enzyme (I-Sce1).  Any cells that convert to GFP+ phenotype, will have completed the process of hi-fidelity homologous recombination.  The readout for HR efficiency the %GFP; however, single cells can be imaged and tracked by time lapse.  This adds dimension of time and analysis of descendants to evaluate and changes in repaired cell behavior. Thus, one major advantage of this kit is that it is operating in the context of a live/living cell. This means that the results will be highly physiological because events are transpiring within the cell in real time.  In addition, because it is florescence-based, it is possible to use single cell imaging to track individual cells that undergo repair and also to track the descendants of those cells using time lapse imaging.

Kit Contents  

1. HR-HeLa Cells frozen in cryoprotection medium. Store at -80^oC for not more than 1 week before thawing.  Alternatively, you may receive a T-25 flask of cells ready for culture. These cells have the DR-HRGFP Gene stably integrated into the genome.  We recommend maintaining these cells in 1 ug/ml puromycin.
2. I-Sce1 gene expression plasmid, pSCE1 (store at -20^o or 4^oC). See tube label for precise concentration of DNA
3. DR-HRGFP(store at -20^o or 4^oC). See tube label for precise concentration of DNA; this plasmid has a puromycin selection gene for counter selection.
4. Puromycin at 2mg/ml (1mL total).
5. The desired quantity of plasmid should be specified in your order.