Dataset Information

Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes.

ABSTRACT: Genetically encoded effectors are important tools for probing cellular function in living animals, but improved methods for directing their expression to specific cell types are required. Here, we introduce a simple, versatile method for achieving cell-type-specific expression of transgenes that leverages the untapped potential of "coding introns" (i.e., introns between coding exons). Our method couples the expression of a transgene to that of a native gene expressed in the cells of interest using intronically inserted "plug-and-play" cassettes (called "Trojan exons") that carry a splice acceptor site followed by the coding sequences of T2A peptide and an effector transgene. We demonstrate the efficacy of this approach in Drosophila using lines containing suitable MiMIC (Minos-mediated integration cassette) transposons and a palette of Trojan exons capable of expressing a range of commonly used transcription factors. We also introduce an exchangeable, MiMIC-like Trojan exon construct that can be targeted to coding introns using the Crispr/Cas system.

SUBMITTER: Diao F

PROVIDER: S-EPMC4373654 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature

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Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes.

Diao Fengqiu F Ironfield Holly H Luan Haojiang H Diao Feici F Shropshire William C WC Ewer John J Marr Elizabeth E Potter Christopher J CJ Landgraf Matthias M White Benjamin H BH

Cell reports 20150226 8

Genetically encoded effectors are important tools for probing cellular function in living animals, but improved methods for directing their expression to specific cell types are required. Here, we introduce a simple, versatile method for achieving cell-type-specific expression of transgenes that leverages the untapped potential of "coding introns" (i.e., introns between coding exons). Our method couples the expression of a transgene to that of a native gene expressed in the cells of interest usi ...[more]

PMID: 25732830

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Project description:Quantitative susceptibility mapping employs regularization to reduce artifacts, yet many recent denoisers are unavailable for reconstruction. We developed a plug-and-play approach to QSM reconstruction (PnP QSM) and show its flexibility using several patch-based denoisers. We developed PnP QSM using alternating direction method of multiplier framework and applied collaborative filtering denoisers. We apply the technique to the 2016 QSM Challenge and in 10 glioblastoma multiforme datasets. We compared its performance with four published QSM techniques and a multi-orientation QSM method. We analyzed magnetic susceptibility accuracy using brain region-of-interest measurements, and image quality using global error metrics. Reconstructions on glioblastoma data were analyzed using ranked and semiquantitative image grading by three neuroradiologist observers to assess image quality (IQ) and sharpness (IS). PnP-BM4D QSM showed good correlation (β = 0.84, R2 = 0.98, p < 0.05) with COSMOS and no significant bias (bias = 0.007 ± 0.012). PnP-BM4D QSM achieved excellent quality when assessed using structural similarity index metric (SSIM = 0.860), high frequency error norm (HFEN = 58.5), cross correlation (CC = 0.804), and mutual information (MI = 0.475) and also maintained good conspicuity of fine features. In glioblastoma datasets, PnP-BM4D QSM showed higher performance (IQGrade = 2.4 ± 0.4, ISGrade = 2.7 ± 0.3, IQRank = 3.7 ± 0.3, ISRank = 3.9 ± 0.3) compared to MEDI (IQGrade = 2.1 ± 0.5, ISGrade = 2.1 ± 0.6, IQRank = 2.4 ± 0.6, ISRank = 2.9 ± 0.2) and FANSI-TGV (IQGrade = 2.2 ± 0.6, ISGrade = 2.1 ± 0.6, IQRank = 2.7 ± 0.3, ISRank = 2.2 ± 0.2). We illustrated the modularity of PnP QSM by interchanging two additional patch-based denoisers. PnP QSM reconstruction was feasible, and its flexibility was shown using several patch-based denoisers. This technique may allow rapid prototyping and validation of new denoisers for QSM reconstruction for an array of useful clinical applications.

Dataset Information

Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes.

Publications

Plug-and-play genetic access to drosophila cell types using exchangeable exon cassettes.

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