Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.
PLoS One. 2013 Jul 8;8(7):e70339. doi: 10.1371/journal.pone.0070339. Print 2013.
The development of forward genetic screens in human haploid cells has the potential to transform our understanding of the genetic basis of cellular processes unique to man. So far, this approach has been limited mostly to the identification of genes that mediate cell death in response to a lethal agent, likely due to the ease with which this phenotype can be observed. Here, we perform the first reporter screen in the near-haploid KBM7 cell line to identify constitutive inhibitors of NF-κB. CYLD was the only currently known negative regulator of NF-κB to be identified, thus uniquely distinguishing this gene. Also identified were three genes with no previous known connection to NF-κB. Our results demonstrate that reporter screens in haploid human cells can be applied to investigate the many complex signaling pathways that converge upon transcription factors.
在人类单倍体细胞中进行正向遗传筛选有可能改变我们对人类特有的细胞过程的遗传基础的理解。到目前为止，这种方法主要局限于识别介导细胞死亡的基因，这可能是因为这种表型很容易被观察到。在这里，我们在近单倍体 KBM7 细胞系中进行了第一次报告基因筛选，以鉴定 NF-κB 的组成性抑制剂。CYLD 是唯一被鉴定出的 NF-κB 的已知负调节剂，因此这个基因具有独特性。还鉴定出了三个以前与 NF-κB 没有已知联系的基因。我们的结果表明，在人类单倍体细胞中的报告基因筛选可以应用于研究许多复杂的信号通路，这些信号通路汇聚到转录因子上。
