An efficient method for generating human somatic cell gene knockouts.
An efficient method for generating human somatic cell gene knockouts.
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DOI：
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发表时间：
2004-02
影响因子：
1.4
通讯作者：
Manu Kohli;C. Rago
中科院分区：
文献类型：
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作者：
Manu Kohli;C. Rago
浸提液：人类基因组计划绘制了一幅地图，详细描述了一个巨大的遗传前沿，将继续为人类疾病的治疗提供有用的见解。大量的未表征基因反映了我们的进步程度和机会的财富。功能基因组学将广泛影响我们对疾病的理解，并照亮更好的治疗方法。确定基因功能的最确定的方法之一是通过敲除方法特异性地敲除基因，从而允许基因匹配（即，同基因的）敲除和野生型对照。基因敲除技术已经在多种模式生物中进行，包括细菌、酵母、鸡和啮齿动物。虽然这些研究可能有助于推断人类基因功能，但很明显，同源物在功能上并不总是相同的。研究人类细胞中基因功能的最佳方法之一是产生人类体细胞基因敲除。然而，这种方法在历史上是低效的，导致采用反义或RNA干扰（siRNA，短干扰RNA）技术的“敲除”方法的广泛使用。这些方法减少而不是消除特定基因的表达，并且通常具有使基因功能分析复杂化的非特异性效应。
Extract: The Human Genome Project has produced a map detailing a vast genetic frontier that will continue to provide useful insights for the treatment of human diseases. The large number of uncharacterized genes reflects the degree of our progress and the wealth of opportunity. Functional genomics will broadly impact our understanding of disease and illuminate the path to better therapeutics. One of the most definitive ways to determine gene function is to specifically inactivate a gene through knockout approaches, thereby permitting comparisons between genetically matched (i.e., isogenic) knockout and wild-type controls. Gene knockout technologies have been performed in a variety of model organisms, including bacteria, yeast, chickens, and rodents. Though these studies might be useful for inferring human gene function, it is clear that homologues are not always functionally identical. One of the best ways to study gene function in human cells is to generate a human somatic cell gene knockout. However, this approach has historically been inefficient, resulting in the widespread use of "knockdown" approaches employing antisense or RNA interference (siRNA, short interfering RNA) technologies. These approaches reduce, rather than eliminate, the expression of a particular gene and often have non-specific effects that complicate the analysis of gene function.