Targeted gene alteration in Caenorhabditis elegans by gene conversion

Barrett, Peter L; Fleming, John T.; Göbel, Verena

doi:10.1038/ng1459

Cited by 37 publications

(38 citation statements)

References 35 publications

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“…However, semistable, transgenic lines with extrachromosomal arrays containing many copies of injected DNA are relatively simple to generate (Mello et al 1991) and provide templates for DSB repair after excision of the endogenous Tc1 transposon (Plasterk and Groenen 1992). The first examples of endogenous gene editing, including tagging a gene with GFP, were based on Tc1 excision (Barrett et al 2004). This strategy has the disadvantage that Tc1 is active only in mutator strains.…”

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confidence: 99%

Exciting Prospects for Precise Engineering of Caenorhabditis elegans Genomes with CRISPR/Cas9

Frøkjær-Jensen

2013

Genetics

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With remarkable speed, the CRISPR-Cas9 nuclease has become the genome-editing tool of choice for essentially all genetically tractable organisms. Targeting specific DNA sequences is conceptually simple because the Cas9 nuclease can be guided by a single, short RNA (sgRNA) to introduce double-strand DNA breaks (DSBs) at precise locations. Here I contrast and highlight protocols recently developed by eight different research groups, six of which are published in GENETICS, to modify the Caenorhabditis elegans genome using CRISPR/Cas9. This reverse engineering tool levels the playing field for experimental geneticists.Progress in science depends on new techniques, new discoveries and new ideas, probably in that order.Sydney Brenner (Robertson 1980, from the symposium

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confidence: 99%

Exciting Prospects for Precise Engineering of Caenorhabditis elegans Genomes with CRISPR/Cas9

Frøkjær-Jensen

2013

Genetics

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“…DNA damage-related neurotic procedures including carcinogenesis [229][230][231][232], maturing [233][234][235][236][237][238][239], and neurodegenerative illnesses are additionally ranges of dynamic research in C. elegans. This exploration has both set up the significance of C. elegans as a model for the investigation of genotoxic specialists (because of protection of the DNA harm reaction) and massively expanded its utility in such reviews by giving an abundance of corresponding and relevant natural data identified with the obsessive reactions to DNA harm in this living being.…”

Section: Pathwaysmentioning

confidence: 99%

Discovering Capacity in Novel Targets: C. Elegans as a Model Life Form in Toxicological Research

Ac¹,

Costa²,

Tec³

et al. 2017

J Clinic Toxicol

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The nematode Caenorhabditis elegans has risen as a critical creature show in different fields including neurobiology, formative science, and hereditary qualities. Qualities of this creature demonstrate that have added to its prosperity incorporate its hereditary manipulability, invariant and completely depicted formative program, all around portrayed genome, simplicity of support, short and productive life cycle, and little body estimate. These same elements have prompted to an expanding utilization of C. elegans in toxicology, both for robotic reviews and highthroughput screening approaches. We depict a portion of the exploration that has been completed in the territories of neurotoxicology, hereditary toxicology, and ecological toxicology, and additionally high-throughput tries different things with C. elegans including all inclusive screening for sub-atomic focuses of harmfulness and fast danger evaluation for new chemicals. We contend for an expanded part for C. elegans in supplementing other model frameworks in toxicological research.

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“…However, the frequency of recombination between the genome and the homologous transgene was too low to be used as an efficient technique for genome engineering. This strategy was reinvestigated using more efficient mutator backgrounds (Barrett et al 2004). Results indicated that it was possible to generate custom deletions and insert a gfp sequence in a locus.…”

Section: Custom Knockout and Knock In Alleles By Transgene-instructedmentioning

confidence: 99%

Manipulating the Caenorhabditis elegans genome using mariner transposons

Robert

Bessereau

2009

Genetica

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Tc1, one of the founding members of the Tc1/mariner transposon superfamily, was identified in the nematode Caenorhabditis elegans more than 25 years ago. Over the years, Tc1 and other endogenous mariner transposons became valuable tools for mutagenesis and targeted gene inactivation in C. elegans. However, transposition is naturally repressed in the C. elegans germline by an RNAi-like mechanism, necessitating the use of mutant strains in which transposition was globally derepressed, which causes drawbacks such as uncontrolled proliferation of the transposons in the genome and accumulation of background mutations. The more recent mobilization of the Drosophila mariner transposon Mos1 in the C. elegans germline circumvented the problems inherent to endogenous transposons. Mos1 transposition strictly depends on the expression of the Mos transposase, which can be controlled in the germline using inducible promoters. First, Mos1 can be used for insertional mutagenesis. The mobilization of Mos1 copies present on an extrachromosomal array results in the generation of a small number of Mos1 genomic insertions that can be rapidly cloned by inverse PCR. Second, Mos1 insertions can be used for genome engineering. Triggering the excision of a genomic Mos1 insertion causes a chromosomal break, which can be repaired by transgene-instructed gene conversion. This process is used to introduce specific changes in a given gene, such as point mutations, deletions or insertions of a tag, and to create single-copy transgenes.

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Targeted gene alteration in Caenorhabditis elegans by gene conversion

Cited by 37 publications

References 35 publications

Exciting Prospects for Precise Engineering of Caenorhabditis elegans Genomes with CRISPR/Cas9

Exciting Prospects for Precise Engineering of Caenorhabditis elegans Genomes with CRISPR/Cas9

Discovering Capacity in Novel Targets: C. Elegans as a Model Life Form in Toxicological Research

Manipulating the Caenorhabditis elegans genome using mariner transposons

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