...alternative method for generating transgenic mice strains and for non-invasive in vivo gene editing. The bottom...repression, transgenic mouse line creation, and in vivo gene editing of mice, with the potential to be used for... Primary cells are often refractory to gene delivery methods, but the researchers was able to edit hiPSCs... stimulation, respectively. Primary cell type Gene targeted by guide Editing efficiency hiPSCs  EMX1...cells when using a gRNA targeting the Fto gene, while editing was not detected in cells electroporated with... used Nanoblades to disrupt the tyrosinase (Tyr) gene in mouse zygotes, which results in albinism in mice....  Nanoblades can also be used to non-invasively gene edit the liver of adult mice. When mice were IV ...

...toolkits available at Addgene.  C. elegans open resources The C. elegans Neuronal Gene Expression Map & Network...expression of your gene of interest.  SapTrap Tool Kit for CRISPR Cas9 edits At Addgene, we have the entire...-throughput CRISPR/Cas9 gene modification in Caenorhabditis elegans." Genetics 202.4 (2016): 1277-1288... The CeNGEN goal is to establish a comprehensive gene expression atlas of every C.elegans  neuron. Scientists... C. elegans embryo when all essential embryonic genes are individually depleted via RNA interference (...based bipartite expression system that can regulate gene expression in C.elegans. In this system a promoter...that can bind to activation sequences upstream of genes to stimulate expression (Wang et al., 2016). In ...

...Plasmids designed to express genes in a given host cell type are generally broken down into two broad categories...discussed the promoter's role in the initiation step of gene transcription; today we'll provide an overview on...define the end of a transcriptional unit (such as a gene) and initiate the process of releasing the newly...machinery. Terminators are found downstream of the gene to be transcribed, and typically occur directly ...studies focus on promoter strength as a determinant of gene expression levels, the terminator also plays an ... to variability in RNA half-life, and ultimately gene-expression. Polyadenylation is the post-transcriptional... in a Promoter-Dependent Manner. Hum Gene Ther. 2008. PMID: 18627247 Zufferey R, Donello JE, Trono D, ...

...Repair is Used to Edit Genes Read How Non-Homologous End Joining is Used to Delete Genes Additional Resources... CRISPR gene therapy, it’s important to realize that DMD is a simpler case than other genetic diseases...there is still no good treatment. The dystrophin gene is very large (79 exons), but much of the sequence...Dystrophin’s size makes it difficult to deliver via gene therapy, so researchers have set their sights on... cells expressed functional dystrophin. Editing genes in vivo is of course much more difficult than in...cells, where precision editing of the dystrophin gene would take place, with minimal risk of off-target...clinically with CRISPR. Many obstacles remain for CRISPR gene editing, but given the speed at which the technology...

...repression of your target gene, activation and repression of different genes in a single cell, and localization...replication and transcription as well as reporter genes. To learn how you can use minigenomes in your research... them in action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (... (AAVs) are commonly used to deliver a genes of interest to a variety of cell types in-vivo. These are...localization of a fluorescent signal to your target gene. Read this post to find out more about how the Casilio...are widely used by the optogenetics and chemogenetics communities to activate neurons and neuronal pathways...guest writers. Your contributions make Addgene and the Addgene Blog useful resources for the biological...

...through this challenging sequence. Transgene The transgene is the gene that will be delivered by the AAV..., or your gene of interest. Promoter The promoter drives the expression of the AAV transgene. On a plasmid...plasmid which contains the structural and packaging genes, the adenoviral helper plasmid which contains the...plasmid map, promoters are upstream or 5’ of the gene they control. Promoters can be organism or tissue...for the successful and specific expression of your gene of interest. References Haery L, Deverman BE, ...Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods 30:206–213 . https://doi.org/10.1089...control transgene expression. Lox sites are directional 34 bp sequences that can flank the transgene and ...

...fluorescent proteins. Selection using resistance genes usually involves killing the untransformed seedlings...individual. For example, with CRISPR-Cas9 gene editing, transformation may be identified by resistance ...continued production of new alleles or off-target gene editing. Antibiotic selection would only then kill...requires specialized equipment. Reporters for studying gene expression have similar drawbacks, often requiring...Res. (2020).   I can do this by expressing a gene that rescues the mutations, using RUBY as a selection.... et al. A reporter for noninvasively monitoring gene expression and plant transformation. Hortic Res ...presence of a transgene instead, which can’t be done if your selection method kills the transgene free individual...

...cross-cassette interactions in gene expression, and to generate multi-gene knockouts. Mammalian and fly ... to generate anything from individual transcription units (TUs) used to express your favorite gene-of-...While a generic MoClo system for any given species can be repurposed for CRISPR applications (gene insert...DNA assembly tool used to build single- and multi-gene constructs for a variety of applications. MoClo ...several additional capabilities, including conditional gene expression, multiplexed CRISPR, and chromosomal ...). Those looking to express high levels of their gene of interest in yeast should also take a look at ...elements for S. cerevisiae chromosomal integration and gene expression, the system includes a toolkit to express...

...Cre-dependent gene of interest in neurons. This ability to be paired with any Cre-controlled gene of interest...neuroscientists because they efficiently deliver genes to various brain regions. You can partially control...occasionally you may want to further control where your gene of interest is expressed.  A cell-type specific ...and what do they mean? AAV: A Versatile Tool for Gene Expression in Mammals Subscribe to Viral Vector ...! To find more viral vectors available through Addgene, be sure to check out our viral service page and...Vector Blog Posts Additional Resources on Addgene.org Check out our Viral Service Page Browse Additional ...

... allowing for the gene on the toehold switch to be expressed. In this case, the gene encodes the LacZ𝛼...interfere with other genetic pathways making it difficult to link a phenotype to the gene being targeted via...need for mutant genetic backgrounds. The standard RNAi vector, L4440, places a target gene between two inverted..., highly efficient RNAi construct for C. elegans gene knockdown Article contributed by Alyssa Cecchetelli...extensively used in C. elegans research to study gene function. RNAi is commonly achieved by feeding E... from each promoter progresses past the specific gene producing long unspecific RNA fragments that are... most scientists use mutant animals that are genetically altered to overexpress endogenous RNAi pathways...