Development of a miRNA Sensor by an Inducible CRISPR-Cas9 Construct in Ciona Embryogenesis

Wang, Zhuqing; Sun, Xiaohui; Zhang, Xiaoming; Dong, Bo; Yu, Haiyan

doi:10.1007/s12033-021-00324-9

Cited by 7 publications

(2 citation statements)

References 37 publications

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“…Electroporation was performed as previously described [ 25 ] with several modifications. After fertilization and dechorionation, 300 μL of eggs with fresh seawater were mixed with 80 μL of plasmids (40 μg) and 420 μL of 0.77 M D-mannitol (Sigma Aldrich) and electroporated using a Gene Pulser Xcell System (Bio-Rad, Hercules, CA, USA) in 0.4 cm cuvettes.…”

Section: Methodsmentioning

confidence: 99%

Quantitative Phosphoproteomics Reveals the Requirement of DYRK1-Mediated Phosphorylation of Ion Transport- and Cell Junction-Related Proteins for Notochord Lumenogenesis in Ascidian

Wang

Ouyang

Tan

et al. 2023

Cells

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The dual-specificity tyrosine phosphorylation-regulated kinase (DYRK1) phosphorylates diverse substrates involved in various cellular processes. Here, we found that blocking the kinase activity of DYRK1 inhibited notochord development and lumenogenesis in ascidian Ciona savignyi. By performing phosphoproteomics in conjunction with notochord-specific proteomics, we identified 1065 notochord-specific phosphoproteins that were present during lumen inflation, of which 428 differentially phosphorylated proteins (DPPs) were identified after inhibition of DYRK1 kinase activity. These DPPs were significantly enriched in metal ion transmembrane transporter activity, protein transport and localization, and tight junction. We next analyzed the downregulated phosphoproteins and focused on those belonging to the solute carrier (SLC), Ras-related protein (RAB), and tight junction protein (TJP) families. In vivo phospho-deficient study showed that alanine mutations on the phosphosites of these proteins resulted in defects of lumenogenesis during Ciona notochord development, demonstrating the crucial roles of phosphorylation of transmembrane transport-, vesicle trafficking-, and tight junction-related proteins in lumen formation. Overall, our study provides a valuable data resource for investigating notochord lumenogenesis and uncovers the molecular mechanisms of DYRK1-mediated notochord development and lumen inflation.

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Section: Methodsmentioning

confidence: 99%

Quantitative Phosphoproteomics Reveals the Requirement of DYRK1-Mediated Phosphorylation of Ion Transport- and Cell Junction-Related Proteins for Notochord Lumenogenesis in Ascidian

Wang

Ouyang

Tan

et al. 2023

Cells

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“…MICR is shown to be activated by miRNAs or siRNAs in a highly sensitive and specific manner. When used to activate the expression of red fluorescent protein (RFP), MICR can serve as a sensor to monitor the expression or activity of specific miRNAs in cells and in vivo (Wang et al, 2019(Wang et al, , 2021. Moreover, because miRNA expression is highly specific to cell type and disease status (Huang, 2017;Wang et al, 2016), MICR has the potential to enable cell type specific genome editing or gene regulation for dissecting gene function in specific cells or curing diseases.…”

Section: Microrna Inducible Crispr Platformmentioning

confidence: 99%

Controlling CRISPR‐Cas9 by guide RNA engineering

Wang

et al. 2022

WIREs RNA

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The clustered regularly interspaced short palindromic repeats (CRISPR) system is a product of million years of evolution by microbes to fight against invading genetic materials. Around 10 years ago, scientists started to repurpose the CRISPR as genetic tools by molecular engineering approaches. The guide RNA provides a versatile and unique platform for the innovation to improve and expand the application of CRISPR‐Cas9 system. In this review, we will first introduce the basic sequence and structure of guide RNA and its role during the function of CRISPR‐Cas9. We will then summarize recent progress on the development of various guide RNA engineering strategies. These strategies have been dedicated to improve the performance of CRISPR‐Cas9, to achieve precise spatiotemporal control of CRISPR‐Cas9, and to broaden the application of CRISPR‐Cas9. Finally, we will briefly discuss the uniqueness and advantage of guide RNA‐engineering based systems versus those with engineered Cas9 proteins and speculate potential future directions in guide RNA engineering. This article is categorized under: RNA Methods > RNA Analyses In Vitro and In Silico RNA Methods > RNA Nanotechnology Regulatory RNAs/RNAi/Riboswitches > Regulatory RNAs RNA Interactions with Proteins and Other Molecules > RNA‐Protein Complexes

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Improved Genome Editing in the Ascidian Ciona with CRISPR/Cas9 and TALEN

Sasakura

Horie

2023

Methods in Molecular Biology

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Development of a miRNA Sensor by an Inducible CRISPR-Cas9 Construct in Ciona Embryogenesis

Cited by 7 publications

References 37 publications

Quantitative Phosphoproteomics Reveals the Requirement of DYRK1-Mediated Phosphorylation of Ion Transport- and Cell Junction-Related Proteins for Notochord Lumenogenesis in Ascidian

Quantitative Phosphoproteomics Reveals the Requirement of DYRK1-Mediated Phosphorylation of Ion Transport- and Cell Junction-Related Proteins for Notochord Lumenogenesis in Ascidian

Controlling CRISPR‐Cas9 by guide RNA engineering

Improved Genome Editing in the Ascidian Ciona with CRISPR/Cas9 and TALEN

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