...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...

...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...

...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...

...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 ...

...stably integrate a reporter gene that contains a puromycin resistance gene followed by the target site...that targets your desired gene (whose sequence is also found in the reporter gene). A control RFP expression...donor plasmid that contains a puromycin resistance gene linked to an in-frame H2B-GFP. This visual read ... express RFP while cells containing the reporter gene that was cleaved by Cas9 and repaired via HDR using...unedited cells. In an experiment that targeted the gene NFE2L2, the Madison lab found that H2B-GFP expressing...desired genomic edit.  Find SRIRACCHA plasmids at Addgene GEMCherry2, an improved monomeric fluorescent protein...stem cells.  Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter systems...

...inducible yet permanent gene excision. Essentially, LoxP sites are knocked in flanking a gene of interest (or ... the gene for your protein of interest. CRISPR-Cas9 and a custom gRNA is all you need to generate a frameshift...? Is the regulation of an exogenously introduced gene or over the native locus? Let’s explore each of...Cas9 fused to a transcriptional repressor to your gene of interest. It will inhibit transcription instead...cinch. Stably introducing the coding region of a gene (cDNA) can be done by either plasmid or virus. A...Cas9. The system is led, via a custom gRNA, to your gene of interest to temporarily increase expression. ...of action Check out Addgene's Lentiviral Tet-on and AAVS1 targeted Tet-on transgene vector. Degron tags...

... 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...

...functional groups of genes involved in the execution of a specific regulatory regime (such as genes involved in...consortium-generated datasets with a systematic effort to compile NR- and coregulator-related global gene profiling...remain unknown. Together, NRs govern expression of genes involved in a broad range of reproductive, developmental...efficient and timely activation or repression of gene expression (see www.nursa.org for a curated list... NRs and coregulators, and although the reporter gene has over the years evolved from chloramphenicol ...O'Malley, B. W. (2002). Combinatorial control of gene expression by nuclear receptors and coregulators...funding model in nuclear receptor research, and has generated a set of comprehensive datasets that collectively...

...specific genes involved. Downregulation of tumor suppressors and upregulation of proto-oncogenes ­­­can ...click on a gene of interest will direct the investigator to a list of all available Addgene plasmids he... group of genes/proteins that, when expressed, normally encourage cell survival, growth and proliferation... the health of an organism that both classes of genes function normally to maintain balance – encouraging...Technologies, Carlsbad, CA) that represents 180 genes from the Ras signaling pathway. Its major appeal...of vectors to study the function of any of these genes through several different applications such as fluorescent... in the development of cancer. With hundreds of genes involved in the Ras pathway alone and thousands ...

...harnessed MMEJ to create a new method for CRISPR gene knock-in, termed PITCh (Precise Integration into Target ...than that observed with NHEJ. PITCh: Using MMEJ for gene knock-in Building on the lab’s previous work, Sakuma... should target near the last coding exon of your gene of interest. For ideal use, this gRNA should be ... is if the GFP-Puro will alter expression of the gene it follows. For increased versatility, it would ...would be advantageous to adapt PITCh to insert genes into AAVS1, the “safe harbor locus” of the human genome...another strategy researchers can exploit for CRISPR gene editing. In organisms where HDR is downregulated...Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh...

... and the gene that is responsible is understood. It’s possible to actually silence this gene with RNAi...only 1,700 genes total). Hydra (Celina Juliano) This is a known stem cell and regeneration model. Little... human interaction for all of us studying human genes in test tubes and gels). As a freshman in college... I studied development and evolution of histone genes in Strongylocentrotus purpuratus, the spiny purple...direct tests of cause and effect hypotheses between gene function and phenotypes in diverse plants, animals...frog cell lines to develop robust methods for frog gene editing. They edit in a cell line, then take the...nonetheless - the first example was of a pigmentation gene.   Genus Microtus (Zoe Donaldson) Voles have fascinating...

...systems allow researchers to understand bacterial gene clusters that produce useful natural materials, ...capable of robust expression of high G+C content genes, as validated through monitoring the expression ...human mRNA binding proteins with 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode...pooled library from Junjie Chen's lab targets human genes known or suspected to be involved in DNA damage ...Veening is now available with one sgRNA per operon or gene. The sgRNA pool can also target core operons in ...library uses CRISPR cytosine base editors to disrupt gene function by targeting start codons, splice acceptor...find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources...

...choose what gene to focus on. The gene choice had restrictions. It couldn’t be a Covid gene, a toxin, or...Syntaxin3 gene was the winner. We used the next 4-5 weeks to work on cloning the Syntaxin3 gene in a pLenti...after discussion, we sent it for sequencing. The gene was successfully sequenced, confirming that we had...with Amy C., Addgene’s intern coordinator shortly after hearing her talk about Addgene’s internship opportunity... past six weeks. Through Addgene, I learned the basics of lab safety, general work skills — such as using...environment changes them. Addgene gave me the chance to become a real life scientist. Addgene gave me the chance...knowledge more than ever before. Before joining Addgene, I had spent most of my junior year stressed out...

...courses on genetic manipulation more hands on While teaching courses about gene therapy and genetic manipulation...that similar types of thinking go into developing gene therapy strategies for various diseases. That’s ...Inserts, Tags, Origins of Replication, Resistance gene and Selection Markers) and one “Transfer Plasmid...had to pick the best strategy and propose a final gene therapy approach for the disease to be openly discussed...impression was that they had clearer ideas about gene therapy in the following lessons. And let me tell...teach gamma-retrovirus or lentivirus-based ex-vivo gene therapy’ strategies to undergraduates, however you...problem is…how can you make a “hands-on” lesson on genetic manipulation of human stem cells when students ...

... plasmids to generate deletions of candidate genes in euploid iPSCs. “The BeHEARD Addgene reagents will... lentiviral vectors from Addgene, the lab will express these candidate genes in TS iPSCs to identify whether...CTD, which arises due to mutations in the SLC6A8 gene, also known as the creatine transporter. The Association...Congratulations Mark Mellett! Siddharth Prakash: X chromosome genes involved in Turner syndrome Turner syndrome (TS)...can lead to cardiac developmental defects. Eight genes from the X chromosome are expressed at significantly...will facilitate our aims to correct X chromosome gene dosage in TS cells as a first step to reverse the...Germline mutations in one of these parts, the GNB1 gene, which codes for G𝛃1 subunits of G protein complexes...

...comparing gene activation when enhancers were active or silent, they could identify the target genes of previously...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...updated Jun 24, 2020. Epigenetic modifications are an additional layer of control over gene expression that ...proteins that help package DNA, were associated with gene expression or silencing. To alter DNA modifications...oxidation and repair factors that allow for increased gene activation at the targeted site compared to other...the only CRISPR-based technology designed to alter gene expression. Fusing dCas9 to a transcriptional activator...a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers. Nature Biotechnology...