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NATF (Native and Tissue-Specific Fluorescence): A Strategy for Bright, Tissue-Specific GFP Labeling of Native Proteins in .

He S, Cuentas-Condori A, Miller DM
Genetics. 2019 212 (2): 387-395

PMID: 30952669 · PMCID: PMC6553825 · DOI:10.1534/genetics.119.302063

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 of tissues that may obscure cell-specific localization. To overcome these problems, we engineered a Native And Tissue-specific Fluorescence (NATF) strategy that combines genome editing and split-GFP to yield bright, cell-specific protein labeling. We use clustered regularly interspaced short palindromic repeats CRISPR/Cas9 to insert a tandem array of seven copies of the GFP11 β-strand ( ) at the genomic locus of each target protein. The resultant knock-in strain is then crossed with separate reporter lines that express the complementing split-GFP fragment () 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 the receptor auxiliary protein LEV-10 at cell-specific synaptic domains in the nervous system.

Copyright © 2019 by the Genetics Society of America.

MeSH Terms (9)

Animals Caenorhabditis elegans Caenorhabditis elegans Proteins CRISPR-Cas Systems Fluorescence Gene Editing Green Fluorescent Proteins Membrane Proteins Nerve Tissue Proteins

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