CRISPR-mediated Tagging with BirA Allows Proximity Labeling in Toxoplasma gondii

被引:20
作者
Long, Shaojun [1 ]
Brown, Kevin M. [1 ]
Sibley, L. David [1 ]
机构
[1] Washington Univ, Sch Med, Dept Mol Microbiol, St Louis, MO 63110 USA
关键词
Mass spectrometry; Protein interaction network; Proximity labeling; CRISPR; BioID; Proximity ligation;
D O I
10.21769/BioProtoc.2768
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Defining protein interaction networks can provide key insights into how protein complexes govern complex biological problems. Here we define a method for proximity based labeling using permissive biotin ligase to define protein networks in the intracellular parasite Toxoplasma gondii. When combined with CRISPR/Cas9 based tagging, this method provides a robust approach to defining protein networks. This approach detects interaction within intact cells, it is applicable to both soluble and insoluble components, including large proteins complexes that interact with the cytoskeleton and unique microtubule organizing center that comprises the apical complex in apicomplexan parasites.
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页数:17
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