We narrowed to 932 results for: Gene

... you started. Any given gene’s webpage will have information on where the gene is expressed within the...from mutations in the gene, and whether any publications have mentioned the gene. The Bloomington Drosophila...also has collections of flies for in vivo gene knock down and gene expression. In the next post, you’ll ...favorite gene (YFG) in Drosophila (and even if there’s not!) the wealth of Drosophila genetic tools available...PhD student at Harvard University. Do you have a gene of interest but have run into a wall trying to study...it? It happens. Is it an evolutionarily conserved gene? Can you find an ortholog in the Drosophila genome...a chance of seeing a phenotype since additional genes that can do the same function are less likely to...

... cassette to the gene of interest. The gene to be expressed is amplified using a gene-specific forward...sgRNA-targeted gene, resulting in increased activation of endogenous transcription of the target gene. The brilliant...of downstream genes. Nihongaki et al describe the targeted activation of endogenous genes and detail the...efficiency and gene expression. The pUCXKT vector ensures that all screened plasmids contain a gene variant*...resistance gene (missing the first two codons). Two stop codons are inserted between the truncated gene and ... inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (...on target gene activation. The novel dCas9-VPR activator is capable of enhancing target gene expression...

...several candidate inhibitors of gene X in a screen. You generate a gene X knockout in a cancerous background...inhibit, activate, or knockout many genes at a time to identify ‘hits’ - genes that affect disease progression...some of the candidate genes! Disease model generation CRISPR has made the generation of genome edits accessible.... These hits can reveal individual genes or sometimes even genetic pathways that are worth investigating...known – repeat expansion, gene knock out, etc. – CRISPR can be used to generate cell line or animal models...approaches. The first route involves identifying a gene/pathway to develop a drug for (aka a druggable target...straightforward causes (a specific mutation in a known gene = disease onset) may be best served by this type...

... popular tool for gene delivery, but it has a drawback: how do you ensure your gene goes where you want...cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved sequences...projections. Generating animal models: Transgenic animals allow for the expression of genes in particular...Additional resources on the Addgene blog Read more about using AAV for retinal gene therapy Find more about...want it to? Knowing that a gene is expressed in a particular cell type is important not only for translational...translational research, such as gene therapy, but also basic research. To improve cell-type specificity of... transcription initiation sites of at least two genes with high expression and high cell-type specificity...

... after light stimulation. Under those optogenetic controls, gene expression levels reliably increased ...could be by applying it to altering gene expression via epigenetics with the addition of histone modifiers...conventional optogenetic tools." In total, the researchers applied the LITE system to 28 different genes to establish...transcriptional effectors (LITEs) designed to bind specific genes and turn them on or off, literally at the flip of...are able to react and learn. All of this requires genes to be regulated dynamically.” To understand how ... tools to manipulate the expression of specific genes in the brain in reversible fashion. And that’s exactly...demonstrated use of the LITEs to control gene expression in mouse neurons and in the brains of living mice ...

...you are trying to do: gene knockout, a specific base edit, or modulation of gene expression. Location ...Created with BioRender.com. The hammer: Gene knockout by NHEJ Gene knockout with CRISPR technology is usually...multiple sequences targeting a gene to score in order to call that gene a hit.  This is an exciting time...optimize numerous parameters to generate the desired edit. The wrench: Gene activation and inhibition by ...edits and to perform functional screens to identify genes involved in a phenotype of interest. This blog post...indels, it's not so important what location in the gene you target, but it is important that your gRNA sequence...creating a non-functional allele. For a 1 kilobase gene, since potential target sites occur ~1 in every ...

...acid sequence from within Snapgene, we can confirm the identity of the gene and check for any mutations...we identify the gene, we can easily annotate it by making a custom feature in Snapgene. This saves us ...frames, which is often our first clue about where the gene of interest might be located. By easily using BLAST...plasmids. We’ve also talked about how our new Snapgene generated maps provide improved feature detection with...’ll walk you through how a scientist at Addgene uses Snapgene to confirm the sequence of a plasmid and...through our Snapgene powered maps and sequence analysis tools.     The main ways we use SnapGene for plasmid...version of Snapgene to visualize the sequence and look for features. The first thing that the Snapgene software...

...Strategies to Make Gene Therapies More General (more than correcting a defect in a single gene) 26:00-28:39 ...39 : Methods for Delivering Multiple Genes to Cells for Gene Therapy 28:40-35:10 : GFP Activatable Tools...Targeting in Retinal Gene Therapies 10:17-14:34 : Progress in Treating Macular Degeneration and Other Retinopathies...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed... gene therapy...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     References...plasmids from this publication at Addgene. Resources on the Addgene Blog Read our Recent Blog Post on...

... epitope tagging of a large cohort of genes (e.g. tagging genes representing a protein super family). ...knock-in for 96 different genes in a single go (Figure 3). We were able to tag >30 genes/plate in the first ...designed for ~250 genes, we have had about 30% tagging success, although many of these genes were expressed...facilitated easy generation of clonal knock-in lines that we now use for IP and ChIP assays. Tag 96 genes at a time...Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian .... Additional Resources on the Addgene Blog Plasmids for endogenous gene tagging in human cells Plasmids...Fluorescent tagging of endogenous genes with SapTrap Resources on Addgene.org Browse CRISPR plasmids Check...

... with ITRs that flank the gene cassette, which contains a promoter, the gene of interest, and a polyA ... DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked by two ITRs...vectors, modifications can affect gene transfer efficiency and transgene expression (Zhou et al. 2017). ...Terminal Repeat Sequence Role in Vector Transgene Expression. Hum Gene Ther, 31(3–4), 151–162. https://doi.org...biological tool that allow scientists to express a gene of interest in a cell or animal model. AAV plasmids...ITRs, and the ITRs are retained with the gene in a recombinant AAV (rAAV) vector (Figure 1). Everything...plasmids. In an AAV vector plasmid, they flank the gene of interest. A single ITR consists of palindromic...

...an unpublished set of Zika gene plasmids to Addgene’s repository. The genes, from the Zika virus Asian...vectors targeting their genes of interest to generate mAID fusions to endogenous genes. Cell lines with the...targeting a total of 20,121 genes within the human genome (the identity of the genes and target sequences can...expression level for their gene of interest. Although the parts involved in altering gene expression are relatively...contains ~90,000 gRNAs that target ~15,000 genes with 6 gRNAs per gene. The second is a "supplementary" library...library to generate a population of HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA... greatly simplified the process of generating multiple E. coli gene expression plasmids with the Cross-disciplinary...

...been used for many years for in vivo delivery and gene therapy and represent one of the most studied viruses...CRISPR-Cas. Figure 1: Schematic of targeted gene knock-out (left) and targeted DNA knock-in (right...binds to target DNA. In the final step of targeted gene knock-out (left), a fragment is deleted from two... This capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral...Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an insertion, a deletion or a frameshift...with Cas9 and a gRNA designed to target a mutated gene. In a Scientific Reports paper introducing the delivery... CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various cell types...

...efforts to regulate gene expression were limited to inducible promoters or direct gene knockout. In contrast... of gene expression. They successfully knock down expression of essential and non essential genes in a.... These could be genes, specific polymorphisms, including antibiotic resistance genes. This approach successfully...resistance pCRISPR: carries a spacer targeting the gene of interest and kanamycin resistance E. coli carrying...wild-type sequence persists, Cas9 will cleave the gene of interest, inducing a lethal DSB.     This system.... pCas contains Cas9 and the phage recombination gene lambda Red. pTargetF contains the specific gRNA(... cleavage induces HDR to directly modify a given gene. CRISPR-tranposons By combining CRISPR editing and...

...Adeno-Associated Viral Vector-Mediated Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https...Specificity of gfaABC1D-Driven Transgene Expression in AAV-Mediated Gene Transfer. Molecular Therapy - ...easier to do the validation first.  AAV-mediated gene expression can differ extensively depending on the... cells in far away brain regions expressing your gene of interest. This can cause issues in tracing studies...can occur. Promoters The promoter controls how the gene of interest will be expressed; it can be ubiquitous...chicken-beta-actin or CAG can drive strong early gene expression, but can become methylated over time,...tyrosine-hydroxylase (TH), can limit which cells express your gene. Off-target expression can still occur (Fig. 3) ...

...this plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY...already have a pBAD33-based plasmid expressing your gene of interest, all you have to do is co-transform ...co-transform pBLADEONLY. That way, a previously cloned gene of interest, under the pBAD promoter, can be controlled... Resources on Addgene.org Check out the Molecular Biology Reference Download Addgene's Plasmids 101 eBook...simply more precise. “We show that you have less heterogeneity in the population and you can do all of these...permission.   BLADE is just one of the several optogenetic tools that Di Ventura’s lab and Khammash's lab...lab have created and shared with Addgene. You can find all the plasmids deposited from Di Ventura's lab...

...using AAV as a tool for gene expression in mammals. Resources on Addgene.org Search for AAV vectors ...One of the biggest barriers to gene therapy treatment of central nervous system (CNS) diseases is the...has not been identified. While LY6A or other Ly6 gene family members are linked to viral susceptibility...large phenotypic effects such as those that affected gene expression, mRNA splicing, or protein coding regions...as non-permissive to PHP.B CNS transduction. Two genes were associated with the PHP.B permissive phenotype...permissiveness. The Wilson Lab also looked for the gene variant responsible for the PHP.B permissive phenotype...inheritance pattern that suggested there was a single gene that had two codominant versions or alleles that...

...cost-effective manipulation of gene expression in the CNS. Molecular Neurodegeneration, 15(1), Article 1. https...system, in which the required adenoviral helper genes are supplied on two different packaging plasmids...pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing replication ...replication factors and serotype-specific capsid genes.  In this helper-free system, HEK293T cells, expressing Ad...transfected with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly...derived inverted terminal repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org...genomes can limit production of recombinant AAV-2. Gene Therapy, 6(7), Article 7. https://doi.org/10.1038...

...while G418 (geneticin) is effective at inhibiting the eukaryotic ribosome. One resistance gene on your vector...used in the lab contain an antibiotic resistance gene to allow for antibiotic selection, but these resistance...aspect of ribosomal function. A puromycin resistance gene on your vector can protect both plasmid-carrying...block protein synthesis, and the neomycin resistance gene protects against any of them. But different aminoglycosides...cause mutations even in cells with the resistance gene, so it isn’t a good fit for all experiments. And...experiment should still include an antibiotic resistance gene for growth in bacteria. Expression You’re almost...systems. But will your cells actually express your gene of interest? Many promoters are species-specific...

...for doing gene targeting, reporter generation, controllable gene knockouts, and inducible gene inactivation...Alec Kimmelman targets mouse metabolic genes with ~6 gRNAs per gene.  CRISPR-Act3.0, developed by Yiping... Shirley Liu's lab targets all mouse genes with 10 gRNAs per gene. The guides are optimized to maximize... to develop CRISPR-based approaches for studying gene function in these systems. As a part of this effort...Yiping Qi's lab, allows simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified...opportunities for investigating genetic mechanisms related to morphogenesis, cell fate specification, and...Lab have compiled a new toolkit for performing genetics studies in organoids. The Organoid EasyTag System...

...the Grousbeck Family Chair in Gene Therapy, Director of the Grousbeck Gene Therapy Center, Associate Director...nonprofit companies. His lab focuses on gene therapy work for genetic diseases and over the past few months...deposited AAV constructs for gene delivery. In 2018, he interviewed Addgene co-founder Melina Fan and former...PhD Khandan Baradaran’s expertise in viral safety, gene therapy vector development, vaccine development,...Dimension Therapeutics, she helped build and grow the gene therapy division of the company.  Ultragenyx’s goal... systematically test the function of all 20,000 genes in disease models and then focus drug discovery ...to crises like those and ended up in the field of gene therapy.” He’s also worked on vaccines throughout...