A genetic toolkit for tagging intronic MiMIC containing genes

Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC is inserted in a codon intron (Nagarkar-Jaiswal et al., 2015). These modified...

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Published in:	eLife Vol. 4
Main Authors:	Nagarkar-Jaiswal, Sonal, DeLuca, Steven Z, Lee, Pei-Tseng, Lin, Wen-Wen, Pan, Hongling, Zuo, Zhongyuan, Lv, Jiangxing, Spradling, Allan C, Bellen, Hugo J
Format:	Journal Article
Language:	English
Published:	England eLife Sciences Publications Ltd 23-06-2015 eLife Sciences Publications, Ltd
Subjects:	Animals Artificial Gene Fusion Cell Biology Chromosomes Deoxyribonucleic acid DNA Drosophila Embryos Gene Targeting - methods Genes Genes, Reporter Genetic Vectors Genomes GFP protein tagging Green Fluorescent Proteins - analysis Green Fluorescent Proteins - genetics Immunoglobulins Immunoprecipitation Insects lgl Medicine Mutagenesis, Insertional Neuroscience Plasmids Proteins Recombinase Recombination, Genetic Research Advance RMCE Staining and Labeling - methods Transposases - metabolism lgl cell biology Drosophila neuroscience GFP protein tagging RMCE D. melanogaster
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Abstract	Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC is inserted in a codon intron (Nagarkar-Jaiswal et al., 2015). These modified genes permit numerous applications including assessment of protein expression pattern, identification of protein interaction partners by immunoprecipitation followed by mass spec, and reversible removal of the tagged protein in any tissue. At present, these conversions remain time and labor-intensive as they require embryos to be injected with plasmid DNA containing the exon tag. In this study, we describe a simple and reliable genetic strategy to tag genes/proteins that contain MiMIC insertions using an integrated exon encoding GFP flanked by FRT sequences. We document the efficiency and tag 60 mostly uncharacterized genes.
AbstractList	Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC is inserted in a codon intron (<xref ref-type="bibr" rid="bib17">Nagarkar-Jaiswal et al., 2015 ). These modified genes permit numerous applications including assessment of protein expression pattern, identification of protein interaction partners by immunoprecipitation followed by mass spec, and reversible removal of the tagged protein in any tissue. At present, these conversions remain time and labor-intensive as they require embryos to be injected with plasmid DNA containing the exon tag. In this study, we describe a simple and reliable genetic strategy to tag genes/proteins that contain MiMIC insertions using an integrated exon encoding GFP flanked by FRT sequences. We document the efficiency and tag 60 mostly uncharacterized genes. Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC is inserted in a codon intron (Nagarkar-Jaiswal et al., 2015). These modified genes permit numerous applications including assessment of protein expression pattern, identification of protein interaction partners by immunoprecipitation followed by mass spec, and reversible removal of the tagged protein in any tissue. At present, these conversions remain time and labor-intensive as they require embryos to be injected with plasmid DNA containing the exon tag. In this study, we describe a simple and reliable genetic strategy to tag genes/proteins that contain MiMIC insertions using an integrated exon encoding GFP flanked by FRT sequences. We document the efficiency and tag 60 mostly uncharacterized genes. Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC is inserted in a codon intron ( Nagarkar-Jaiswal et al., 2015 ). These modified genes permit numerous applications including assessment of protein expression pattern, identification of protein interaction partners by immunoprecipitation followed by mass spec, and reversible removal of the tagged protein in any tissue. At present, these conversions remain time and labor-intensive as they require embryos to be injected with plasmid DNA containing the exon tag. In this study, we describe a simple and reliable genetic strategy to tag genes/proteins that contain MiMIC insertions using an integrated exon encoding GFP flanked by FRT sequences. We document the efficiency and tag 60 mostly uncharacterized genes. DOI: http://dx.doi.org/10.7554/eLife.08469.001 Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the generation of a new exon that encodes a protein tag when the MiMIC is inserted in a codon intron (Nagarkar-Jaiswal et al., 2015). These modified genes permit numerous applications including assessment of protein expression pattern, identification of protein interaction partners by immunoprecipitation followed by mass spec, and reversible removal of the tagged protein in any tissue. At present, these conversions remain time and labor-intensive as they require embryos to be injected with plasmid DNA containing the exon tag. In this study, we describe a simple and reliable genetic strategy to tag genes/proteins that contain MiMIC insertions using an integrated exon encoding GFP flanked by FRT sequences. We document the efficiency and tag 60 mostly uncharacterized genes.DOI: http://dx.doi.org/10.7554/eLife.08469.001
Author	Lin, Wen-Wen Lee, Pei-Tseng Pan, Hongling Bellen, Hugo J Lv, Jiangxing Spradling, Allan C DeLuca, Steven Z Nagarkar-Jaiswal, Sonal Zuo, Zhongyuan
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Keywords	lgl cell biology Drosophila neuroscience GFP protein tagging RMCE D. melanogaster
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Snippet	Previously, we described a large collection of Minos-Mediated Integration Cassettes (MiMICs) that contain two phiC31 recombinase target sites and allow the...
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SubjectTerms	Animals Artificial Gene Fusion Cell Biology Chromosomes Deoxyribonucleic acid DNA Drosophila Embryos Gene Targeting - methods Genes Genes, Reporter Genetic Vectors Genomes GFP protein tagging Green Fluorescent Proteins - analysis Green Fluorescent Proteins - genetics Immunoglobulins Immunoprecipitation Insects lgl Medicine Mutagenesis, Insertional Neuroscience Plasmids Proteins Recombinase Recombination, Genetic Research Advance RMCE Staining and Labeling - methods Transposases - metabolism
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Title	A genetic toolkit for tagging intronic MiMIC containing genes
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