Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
CRISPR J. 2022 Feb;5(1):155-164. doi: 10.1089/crispr.2021.0069.
Functional characterization of the multitude of poorly described proteins in the human malarial pathogen, , requires tools to enable genome-scale perturbation studies. Here, we present GeneTargeter (genetargeter.mit.edu), a software tool for automating the design of homology-directed repair donor vectors to achieve gene knockouts, conditional knockdowns, and epitope tagging of genes. We demonstrate GeneTargeter-facilitated genome-scale design of six different types of knockout and conditional knockdown constructs for the genome and validate the computational design process experimentally with successful donor vector assembly and transfection. The software's modular nature accommodates arbitrary destination vectors and allows customizable designs that extend the genome manipulation outcomes attainable in and other organisms.
功能特征的众多描述不佳的蛋白质在人类疟原虫病原体,需要工具来实现基因组规模的干扰研究。在这里,我们提出了 GeneTargeter(genetargeter.mit.edu),一个软件工具,用于自动设计同源重组供体载体实现基因敲除,条件敲低,和表位标记的基因。我们展示 GeneTargeter 促进基因组规模的设计六种不同类型的敲除和条件敲低构建体的基因组和验证计算设计过程实验成功的供体载体组装和转染。软件的模块化性质适应任意目的载体,并允许可定制的设计,扩展基因组操作结果可在和其他生物体。