Robotic high-throughput assay for isolating apoptosis-inducing genes

BioTechniques
Stefan Grimm, Volker Kachel

Abstract

The need to assign functions to genes will increase in upcoming years because of the accumulation of sequence information from various organisms. Here we present a genetic screen for the systematic functional analysis of genes in mammalian cells. The screen is based on high-throughput, transient transfections of individual cDNAs by a robotic system. This construction uses a moveable pipetting head with a novel 96-channel parallel tube-pump array and an integrated de-lidding device. Because of the robot design, the processing speed can reach 2,000 transfections/h. We demonstrate the suitability of the screen to isolate apoptosis-inducing genes as a first read-out. This is achieved in combination with co-transfected reporter plasmids to produce a novel and convenient assay for apoptosis induction.

Citations

Jun 24, 2003·Biological Chemistry·Christian PlankJoseph Rosenecker
Feb 19, 2004·Nature Reviews. Genetics·Stefan Grimm
May 12, 2005·Médecine sciences : M/S·Yoann RoupiozXavier Gidrol
Sep 30, 2004·International Journal of Cancer. Journal International Du Cancer·Kerstin Koenig-HoffmannUlrich Brinkmann
Mar 23, 2005·Nature Methods·Mark IsalanLuis Serrano
Sep 25, 2012·Biochimica Et Biophysica Acta·Stefan Grimm
Feb 18, 2006·BMC Biotechnology·Volker KachelStefan Grimm

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis