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Breakthrough Technology for Functional Genomics in Mammalian Cells

Peking Univeristy, Apr. 11, 2014: One of the most important challenges in the research of life sciences is to attribute physiological functions precisely to responsible genes. Despite the rapid development of toolkits to manipulate individual genes, large-scale screening methods based on the complete loss of gene expression are still missing. A research team led by Prof. Wei Wensheng of the College of Life Sciences, in collaboration with Prof. Huang Yanyi of Biodynamic Optical Imaging Centre (BIOPIC), developed a groundbreaking technique that achieves high-throughput functional genomics in eukaryotes.


Arrayed and pooled screens using RNAi libraries targeting either human or mouse genomes have already been developed and widely used for systematic genetic studies in mammalian cells. However, its limitations are increasingly apparent. In particular, RNAi-based down-regulation of any particular gene expression is not always sufficient to cause phenotypical changes of interest. Taking advantage of the targeted genome editing technologies, Wei lab developed a focused CRISPR/Cas9-based lentiviral library in human cells and a method of gene identification based on functional screening and high-throughput sequencing analysis. Using knockout library screens, they successfully identified the host components essential for the intoxication of cells by two bacterial toxins, which were confirmed by functional validation. The broad application of this powerful genetic screening strategy will not only facilitate the rapid identification of genes important for bacterial toxicity, but will also enable the discovery of genes that participate in a broad range of biological processes.

Given the importance to establish an effective genome-editing technology for functional genomics in a high-throughput fashion, the international competition has been fierce. Two parallel studies reported a similar methodology in Science earlier this year. However, the approach developed from Wei lab is better suited for broader range of cell types, and is particularly advantageous for knowledge-based screening. 

This work was published online on April 9 in Nature(High-throughput Screening of a CRISPR/Cas9 Library for Functional Genomics in Human Cells). The co-first authors are three Ph.D. students from Wei lab, Zhou Yuexin, Zhu Shiyou and Cai Changzu. This work was supported by funds from the National Basic Research Program of China, the National Science Foundation of China, and the Peking-Tsinghua Center for Life Sciences.


Original link: http://www.nature.com/nature/journal/vaop/ncurrent/full/nature13166.html

Edited by: Zhang Jiang

Source: School of Life Sciences