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Rapid generation of hypomorphic mutations.

Nat Commun. 2017; 
Arthur Laura L,Chung Joyce J,Jankirama Preetam,Keefer Kathryn M,Kolotilin Igor,Pavlovic-Djuranovic Slavica,Chalker Douglas L,Grbic Vojislava,Green Rachel,Menassa Rima,True Heather L,Skeath James B,Djuranovic Se
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摘要

Hypomorphic mutations are a valuable tool for both genetic analysis of gene function and for synthetic biology applications. However, current methods to generate hypomorphic mutations are limited to a specific organism, change gene expression unpredictably, or depend on changes in spatial-temporal expression of the targeted gene. Here we present a simple and predictable method to generate hypomorphic mutations in model organisms by targeting translation elongation. Adding consecutive adenosine nucleotides, so-called polyA tracks, to the gene coding sequence of interest will decrease translation elongation efficiency, and in all tested cell cultures and model organisms, this decreases mRNA stabilit... More

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