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- W2893587839 abstract "ABSTRACT GFP labeling by genome editing can reveal the authentic location of a native protein but is frequently hampered by weak GFP signals and broad expression across a range cell types in multicellular animals. To overcome these problems, we engineered a N ative A nd T issue-specific Fluorescence (NATF) strategy which combines CRISPR/Cas-9 and split-GFP to yield bright, cell-specific protein labeling. We use CRISPR/Cas9 to insert a tandem array of seven copies of the GFP11 β-strand ( gfp11 x7 ) at the genomic locus of each target protein. The resultant gfp11 x7 knock-in strain is then crossed with separate reporter lines that express the complementing split-GFP fragment ( gfp1 - 10 ) in specific cell types thus affording tissue-specific labeling of the target protein at its native level. We show that NATF reveals the otherwise undetectable intracellular location of the immunoglobulin protein, OIG-1, and demarcates a receptor auxiliary protein LEV-10 at cell-specific synaptic domains in the C. elegans nervous system." @default.
- W2893587839 created "2018-10-05" @default.
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- W2893587839 date "2018-09-25" @default.
- W2893587839 modified "2023-09-27" @default.
- W2893587839 title "NATF (Native And Tissue-specific Fluorescence): A strategy for bright, tissue-specific GFP labeling of native proteins" @default.
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- W2893587839 doi "https://doi.org/10.1101/426205" @default.
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