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Modifying the C. elegans genome. | Mos1-based methods to engineer the worm genomeMos1-based methods to engineer the worm genome
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Mos1-based methods to engineer the worm genome
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Modifying the C. elegans genome. | Mos1-based methods to engineer the worm genome | wormbuilder.org Reviews
https://wormbuilder.org
Mos1-based methods to engineer the worm genome
wormbuilder.org
MosDEL | Modifying the C. elegans genome.
http://www.wormbuilder.org/mosdel
Darr; Skip to Main Content. Universal MosSCI insertion strains. About Mos1-mediated Deletion (mosDEL). MosDEL is a method to generate targeted deletions in the genome of. The method has the advantages:. Deletions are generated with selection markers. The exact endpoints of a deletion can be specified. Essential genes can be deleted and balanced at the same time. Or NeoR) and fluorescent markers to select against extra-chromosomal arrays. A Mos1 element within 25 kb of the gene/region to be deleted.
Bright MosSCI inserts | Modifying the C. elegans genome.
http://www.wormbuilder.org/test-page/bright-balancers
Darr; Skip to Main Content. Universal MosSCI insertion strains. These insertions are bright enough to be seen on a fluorescence dissection scope and can be useful for moving MosSCI insertions into other genetic backgrounds. OxSi259[Peft-3: GFP cb-unc-119 ] I; unc-119(ed3) III. OxSi257[Peft-3: GFP cb-unc-119 ] I; unc-119(ed3) III. OxSi221[Peft-3: GFP cb-unc-119 ] II; unc-119(ed3) III. Unc-119(ed3) III; oxSi346[Peft-3: GFP cb-unc-119 ] IV. Dpy-13(ox495: cb-unc-119 Pmyo-2: mCherry Punc-122: GFP) IV.
Reagents | Modifying the C. elegans genome.
http://www.wormbuilder.org/reagents
Darr; Skip to Main Content. Universal MosSCI insertion strains. The reagents listed here were made by miniMos insertions into the genome. Some of these may be useful to others and we have therefore deposited many of the strains with the CGC. Some of the uses could be:. Bright miniMos inserts are useful for building double and triple mutants. Bright miniMos inserts can be used to outcross identified mutations. 256x LacO insertions can be used to track chromosome dynamics.
Protocol | Modifying the C. elegans genome.
http://www.wormbuilder.org/minimos/protocol
Darr; Skip to Main Content. Universal MosSCI insertion strains. Make miniMos vector with your transgene. There are vectors with. NeoR, PuroR and HygroR selection or without a selection marker that are compatible with either three-fragment Gateway cloning or restriction/Gibson cloning. See plasmids for info. Maintain healthy injection strain. Healthy worms are much easier to inject and result in higher insertion frequencies. Maintain. Strains on HB101 at 15 C or 20 C. Inject animals and place at 25 C.
Protocol | Modifying the C. elegans genome.
http://www.wormbuilder.org/test-page/protocol
Darr; Skip to Main Content. Universal MosSCI insertion strains. Make targeting vector containing your transgene. Each insertion site has a specific vector and an injection strain that is compatible with that targeting vector. See plasmids and injection strains for info. Maintain injection strain on HB101 bacteria at 20 degrees. Healthy worms are much easier to inject and result in higher insertion frequencies. Maintain. Strains at 15C or 20C. Heat-shock injected animals (optional). Homozygose the inserti...
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How it Works - Cas9-triggered homologous recombination
http://wormcas9hr.weebly.com/how-it-works.html
About the CRISPR/Cas9 system. Cas9 is an endonuclease that functions in bacterial adaptive immunity. In nature, Cas9 binds to two small RNAs: a trans-activating RNA (tracrRNA) that activates its enzymatic activity, and a CRISPR RNA (crRNA) that determines substrate specificity. In a seminal paper. Single guide RNA (sgRNA). Thus, by changing the targeting sequence at the 5' end of the sgRNA, Cas9 can be programmed to target a wide variety of sequences. About Cas9-triggered homologous recombination. That m...
Lanctot Lab - Links
http://www.lanctotlab.org/en/liens_EN.html
WDDD - Worm Dev Dyn Database. EPIC - Expression patterns. Biology of the nucleus. Labs (far from exhaustive list). Int Mouse Strain Resources. Euro Cond Mouse Mutagenesis. Eurexpress (Embryo, in situs). NIH Knockout Mouse Project. Int Gene Trap Consortium. Mutant Mouse Resource NIH. Bioinformatics tools and databases. Genome, protein and plasmid databases. DB of Biological databases. InterPro - Protein Domains. Jaspar (TF Binding sites). KEGG-Kyoto Encyc Genes Genomes. PDB - Protein DataBank. Institute o...
Links and References - Cas9-triggered homologous recombination
http://wormcas9hr.weebly.com/links-and-references.html
Our papers on Cas9-triggered homologous recombination:. Dickinson DJ, Pani AM, Heppert JK, Higgins CD and Goldstein B (2015). Streamlined genome engineering with a self-excising drug selection cassette. Genetics. Early Online. DOI: 10.1534/genetics.115.178335. Dickinson DJ, Ward JD, Reiner DJ and Goldstein B (2013). Engineering the Caenorhabditis elegans. Genome using Cas9-triggered homologous recombination. Nature Methods. 10:1028-1034. DOI: 10.1038/nmeth.2641. Resources for genome-editing techniques:.
Linked List | The Glotzer Lab | Michael Glotzer
http://glotzerlab.uchicago.edu/linked_list.html
The molecular mechanism of cytokinesis. Two cell C. elegans. Embryo microtubules (green) centralspindlin complex (red). Dept of Molecular Genetics and Cell Biology. Program in Cell and Molecular Biology. Weather from Forecast.io. Glotzer Lab Department of Molecular Genetics and Cell Biology.
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Modifying the C. elegans genome. | Mos1-based methods to engineer the worm genome
Darr; Skip to Main Content. Universal MosSCI insertion strains. Fluorescent markers strains with GFP insertions. In collaboration with Ann Rougvie at the CGC, we have generated a set of strains carrying GFP markers at defined genomic locations. The strains are available from the CGC and you can search for the strains based on chromosome, fluorescent marker, and the selection marker (see Fluorescent marker strains. Move to Andrew Fire’s lab at Stanford. Sept 16, 2014. 8216;s lab at Stanford! I will also w...
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