...nearby genes and its role in gene expression, though this may still unexpectedly affect nearby genes (Figure...protein-coding genes thanks to systems like CRISPR/Cas9 that allow them to manipulate full genes and observe..., or permanently disable, protein-coding genes by generating insertions or deletions in the DNA sequence...only 2% of our genome is made of protein-coding genes, and you might be wondering what the rest of our...manipulate and study in comparison to protein-coding genes (Rinn and Chang 2012). Scientists have determined...multiple gRNAs poses the risk of disrupting adjacent genes, especially those regulated by or overlapping with...). The interactions between ncRNAs and adjacent genes are complex and not yet fully understood, so it ...

... 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...

...et al., Adenovirus-mediated gene delivery: Potential applications for gene and cell-based therapies in... AdVs are stripped of regulatory genes E1 and E3. Without these genes, AdVs cannot replicate on their ...et al. "Adenovirus-mediated gene delivery: Potential applications for gene and cell-based therapies in...applications and gene therapy: they can be produced at high titers, can accommodate large transgenes, transduce...recombination-competent adenovirus (RCA) can be generated if the E1 gene from the packaging cell line is transferred...for the generation of RCA and reduced immunogenicity. In addition to E1/E3, non-structural genes E2 and...proteins in trans. The Gutless AdV have generated high interest for gene therapy due their increased cloning...

... 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...

...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...

... 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 ...

...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...