Mouse 3T3 Cell Based Kit for Interrogating HR-DNA Repair using I-Sce1 Site Cleavage

This particular cell-based reporter kit is 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. The kit uses GFP as an in vivo readout for the HR pathway and a plasmid expression vector containing the mega-endonuclease I-Sce1. The host cell is the mouse cell line 3T3 (HR-3T3).  This cell line is easy to handle and will arrest in G1 naturally to allow cell cycle analysis of HR repair events.

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 will find this kit very useful. The 3T3 line is easy to handle and synchronize in G1/S which give the end user additional tools to examine cell cycle arrest behavior and HR Repair.  It is well established that the DNA repair pathway is replete with druggable targets. Agents that target DBNA repair are usually good anti-cancer or anti-viral drugs.  Also, identifying other genes and gene products that interact with DNA repair is important in order to find new drug targets.  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. Mouse 3T3 cells frozen (cyro-protective medium) with the endogenous HR GFP reporter stably integrated. Cells may be sent frozen or ambient.
2. I-Sce1 gene expression plasmid. Customer should specify the Kit size (25, 50, 100 assays corresponding to 50, 100 or 250 ug of I-Sce1 plasmid DNA, respectively).
3. Puromycin Stock.
4. A detailed protocol describing how to set up and use this kit for assaying HR mechanisms in the cell host provided.