Inhibition of no tail (ntl) gene expression in zebrafish by external guide sequence (EGS) technique

Pei, De‐Sheng; Sun, Yuhan; Long, Yong; Zhang, Zhu

doi:10.1007/s11033-007-9063-9

Cited by 12 publications

(10 citation statements)

References 21 publications

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“…2A). In contrast, in the external guide sequence (EGS) technique, the antisense guide is designed to bind to the target mRNA such that together they form a complex of hairpin structures that resemble a tRNA molecule (Pei et al,2007). This structure is then recognized by the endogenous ribonuclease (RNase) P, which then cleaves the mRNA in a manner analogous to the maturation of tRNA from its precursor molecule (Liu and Altman,1996; Rangarajan et al,2004; Pei et al,2007; Fig.…”

Section: Ribozyme‐mediated Strategies For Gene Function Analysismentioning

confidence: 99%

“…Once guide sequence candidates have been identified, they are cloned into the appropriate expression vectors to generate the ribozymes or the external guide sequences. Delivery of these constructs into the embryo is performed following standard injection protocols (Xie et al,1997; Walker et al,2001; Pei et al,2007).…”

Section: Ribozyme‐mediated Strategies For Gene Function Analysismentioning

confidence: 99%

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Current perspectives in zebrafish reverse genetics: Moving forward

Skromne

Prince

2008

Developmental Dynamics

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Use of the zebrafish as a model of vertebrate development and disease has expanded dramatically over the past decade. While many articles have discussed the strengths of zebrafish forward genetics (the phenotypedriven approach), there has been less emphasis on equally important and frequently used reverse genetics (the candidate gene-driven approach). Here we review both current and prospective reverse genetic techniques that are applicable to the zebrafish model. We include discussion of pharmacological approaches, popular gain-of-function and knockdown approaches, and gene targeting strategies. We consider the need for temporal and spatial control over gain/loss of gene function, and discuss available and developing techniques to achieve this end. Our goal is both to reveal the current technical advantages of the zebrafish and to highlight those areas where work is still required to allow this system to be exploited to full advantage. Developmental Dynamics 237:861-882, 2008.

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Section: Ribozyme‐mediated Strategies For Gene Function Analysismentioning

confidence: 99%

Section: Ribozyme‐mediated Strategies For Gene Function Analysismentioning

confidence: 99%

Current perspectives in zebrafish reverse genetics: Moving forward

Skromne

Prince

2008

Developmental Dynamics

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“…Zebrafish has been widely accepted as an excellent model for developmental, toxicological and pharmacological studies to identify conserved developmental signaling pathways and understand mechanisms of human diseases [21]. In this study, we report the cloning of zebrafish abcc1 cDNA, transcriptional expression patterns of this gene in developing embryos and adult tissues, and its roles in detoxification of toxic heavy metals.…”

Section: Introductionmentioning

confidence: 99%

Molecular analysis and heavy metal detoxification of ABCC1/MRP1 in zebrafish

Long

Cui

2010

Mol Biol Rep

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ABCC1/MRP1 belongs to the ATP-binding cassette superfamily and its elevated expression is closely associated with the multidrug resistance of various tumor cells. In normal tissues, ABCC1 confers resistance to a wide variety of xenobiotics and toxicants, demonstrating its important roles in tissue defense. Here, we report the cloning and functional characterization of abcc1 gene in zebrafish. This gene is localized on zebrafish chromosome 3 and contains a 4,557 bp open-reading frame. The deduced polypeptide is composed of 1,518 amino acids, which shares 70% identity with human ABCC1. Phylogenetic analysis revealed that ABCC1 proteins from thirteen vertebrate species are highly conserved during evolution. Transcriptional expression of zebrafish abcc1 gene in developing embryos was examined by whole-mount in situ hybridization and real-time PCR. Transcripts of zebrafish abcc1 gene were detectable in four-cell stage embryos, indicating that this gene is maternally expressed. ABCC1 mRNAs were ubiquitously distributed in embryos before 12 h post-fertilization (hpf) and mainly localized in eyes and brain from 24 to 72 hpf, and in gills from 96 to 120 hpf. In addition, zebrafish abcc1 gene was highly expressed in 1-hpf embryos and detected in all adult tissues examined, with highest expression in testis and lowest in heart and liver. Exposure of ZF4 cells and embryos to CdCl 2 Á2.5H 2 O, HgCl 2 , Pb(NO 3 ) 2 , or Na 3 AsO 4 Á12H 2 O significantly induced transcriptional expression of abcc1 gene. Furthermore, overexpression of abcc1 improved the survival rates of embryos exposed to Cd, Hg or As, while overexpression of a abcc1 mutant (ABCC1-G1420D) sensitized zebrafish embryos to toxic metals. These data indicate that zebrafish ABCC1 has crucial roles in heavy metals detoxification.

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“…Although the utility of EGS-directed cleavage by RNase P had been demonstrated in bacterial, animal, and plant cells (Guerrier-Takada et al 1995;Plehn-Dujowich and Altman 1998;Raj et al 2001;Dreyfus et al 2004;Rangarajan et al 2004;Pei et al 2008), similar possibilities for inhibition of gene expression in yeast cells remained unclear. We show here that M1 3/4EGS RNA alone is efficient in cleavage of yeast target RNA in vitro, and was capable of reducing the target mRNA in vivo by up to 30% as measured by Northern blots.…”

Section: Discussionmentioning

confidence: 99%

“…In several recent studies, the external guide sequence (EGS)-based cleavage by RNase P (Forster and Altman 1990) has been used successfully to inhibit gene expression in bacterial, animal, and plant cells (Guerrier-Takada et al 1997;PlehnDujowich and Altman 1998;Raj et al 2001;Dreyfus et al 2004;Rangarajan et al 2004;Pei et al 2008), while its utility in yeast remains unexplored. Therefore, we undertook this study to test the viability of this approach for disrupting expression of host genes in Saccharomyces cerevisiae.…”

Section: Introductionmentioning

confidence: 99%

Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology

Cheng¹,

Ko²,

Altman³

2011

RNA

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The artificial inhibition of expression of genes in Saccharomyces cerevisiae is not a widespread, useful phenomenon. The external guide sequence (EGS) technology, which is well-proven in bacteria and mammalian cells in tissue culture and in mice, can also be utilized in yeast. The TOP2 and SRG1 genes can be inhibited by~30% with EGSs in vivo. Results in vitro also show convenient cleavage of the relevant transcripts by RNase P and appropriate EGSs. The feasible constructs shown to date have an EGS covalently linked to M1 RNA, the RNA subunit of RNase P from Escherichia coli. Greater efficiency in cleavage of transcripts can be fashioned using more than one EGS targeted to different sites in a transcript and stronger promoters controlling the EGS constructs.

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Inhibition of no tail (ntl) gene expression in zebrafish by external guide sequence (EGS) technique

Cited by 12 publications

References 21 publications

Current perspectives in zebrafish reverse genetics: Moving forward

Current perspectives in zebrafish reverse genetics: Moving forward

Molecular analysis and heavy metal detoxification of ABCC1/MRP1 in zebrafish

Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology

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