Dna transfection into 283T (HEK-293) cells using DreamFect

293T cells were transiently transfected with a 1:1 ratio of plasmids using DreamFect reagent from OZ Biosciences.

This article demonstrate the efficiency of Dreamfect transfection reagent from OZ Biosciences to efficiently deliver plasmid into 293T cells.

article reference: Murfini I, PLoS Genet. 2013 Oct;9(10):e1003910.

Survival of the Replication Checkpoint Deficient Cells Requires MUS81-RAD52 Function.

Murfuni I, Basile G, Subramanyam S, Malacaria E, Bignami M, Spies M, Franchitto A, Pichierri P.

Abstract

In checkpoint-deficient cells, DNA double-strand breaks (DSBs) are produced during replication by the structure-specific endonuclease MUS81. The mechanism underlying MUS81-dependent cleavage, and the effect on chromosome integrity and viability of checkpoint deficient cells is only partly understood, especially in human cells. Here, we show that MUS81-induced DSBs are specifically triggered by CHK1 inhibition in a manner that is unrelated to the loss of RAD51, and does not involve formation of a RAD51 substrate. Indeed, CHK1 deficiency results in the formation of a RAD52-dependent structure that is cleaved by MUS81. Moreover, in CHK1-deficient cells depletion of RAD52, but not of MUS81, rescues chromosome instability observed after replication fork stalling. However, when RAD52 is down-regulated, recovery from replication stress requires MUS81, and loss of both these proteins results in massive cell death that can be suppressed by RAD51 depletion. Our findings reveal a novel RAD52/MUS81-dependent mechanism that promotes cell viability and genome integrity in checkpoint-deficient cells, and disclose the involvement of MUS81 to multiple processes after replication stress.

DreamFect from OZ Biosciences allows transfecting all types of nucleic acids with a very high efficiency; it is fully biodegradable and does not interfere with cellular mechanisms.