We narrowed to 1,147 results for: Gene

...from transient overexpression of protein fusions to gene-edited cells can decrease the number of fluorophores...Fluorescent Protein Should I Use? Resources on Addgene.org Generate Fluorescent Protein Fusions Request Plasmids...in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your... dye ligand (e.g., FlAsH, ReAsH) and a short genetically encoded tetracysteine (Cys4) peptide tag could...TMP-tag) – These widely used systems consist of a genetically encoded enzyme variant tag that reacts specifically...such as paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule...strategies have trade-offs between the size of the genetically encoded tag, the speed and selectivity of the...

...contain either an ampicillin or kanamycin resistance gene. Every step along the way involves good aseptic ...of tube for each plasmid we have at Addgene. Each sample at Addgene has four layers of location which can...pages! Resources Avaliable on the Addgene Blog Read more Inside Addgene Posts! Learn all about Plasmids ...Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the... a microcentrifuge tube. Once a sample enters Addgene’s lab, a series of events is triggered. Each step...fluctuates throughout the year (see graph below). Addgene scientists inspect each sample and make sure they...takes over and prepares the sample for entry into Addgene’s repository. For long-term storage in our facilities...

... Ti plasmids in Agrobacterium tumefaciens. Genes & Genetic Systems 77:1–9 . https://doi.org/10.1266/ggs... Resources on Addgene.org Download the Plasmids 101 eBook  Browse plasmids at Addgene Read our Molecular...plasmid partitioning system in the bacteria, but generally it consists of a centromere-like region on the...Kamruzzaman et al., 2017). Additional resources on the Addgene blog Read all of our Plasmids 101 blog posts Learn...

...likely -/-, 50% are -/+, and 25% are +/+ for your gene of interest. Thus, to find a null clone, you should...edited. For many experiments, the end goal is to generate a clonal cell line, so it’s important to verify... for a needle in a haystack before going on to generate and screen individual clones. If a clonal line...al., NAR). This technique uses the trace file generated in Sanger sequencing and a simple algorithm to...with your sgRNA sequence. The TIDE software will generate a graph representing all insertions and deletions... 4 clones. Did you use a dual guide system to generate your knockout? Then you’re in luck! You don’t ... to visualize an edit. Most dual guide systems generate a deletion large enough that it can be visualized...

...IPTG. T7 RNAP is then available to transcribe the gene of interest from a T7 promoter on the plasmid. Many...::T7 gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein...cytosine within a particular sequence. Browse Addgene's curated list of Bacterial Expression Systems Table...DE3) F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) General protein expression BL21 (DE3) pLysS* Chloramphenicol...ompT lon hsdSB (rB- mB-) gal dcm rne131 (DE3) General expression; not recommended for toxic proteins ...lon hsdSB (rB- mB-) gal dcm araB::T7RNAP-tetA General protein expression  BLR (DE3) Tetracycline RecA-deficient...overexpression? The strains described above should generate sufficient expression levels for most purposes...

...mediated HDR) enables the tagging of endogenous genes close to native expression levels. However, it is...chimera are encoded by DNA. Since researchers can generate almost any DNA sequence in the way that they like...proteins is relatively straightforward. However, generating a fusion while keeping all of the native properties...challenging. In this blog I discuss strategies to generate fusion proteins and highlight some aspects of ... from Aequorea victoria and its variants are genetically encoded fluorescent probes. One of the limitations...restriction enzymes in the multiple cloning site for generating fusions. For the POI-fluorescent protein fusions...sufficient. For the fluorescent protein-POI fusions, we generate straight fusions, without any linker. When both...

...achieved through, for example, a previously induced gene knockout. Besides amine- and DNA-binding dyes, nucleoside... on Addgene.org Ready-to-use recombinant antibodies at Addgene Addgene's Antibody Guide Addgene's Antibody... combines a marker with a signal. For clarity, Addgene uses 'labeling' to refer to antibodies and 'staining...Find streptavidin plasmids and biotin plasmids at Addgene!     Figure 1: Indirect detection of target...signal. Dump gating When you are working with heterogeneous cell mixes, like bulk lympho-/splenocytes, you...Antibody Plasmid Collection Resources on the Addgene blog Antibodies 101: Reading a Flow Plot Antibodies...

...do not naturally express the protein or have had gene expression knocked down or knocked out. If you are...antibodies Resources on Addgene.org Find written and video protocols Visit Addgene's antibodies collection...plasmid encoding your protein of interest from Addgene, transfected it into your target cells, now what...wet, semi-dry, and dry but all follow the same general principle where the gel and membrane are “sandwiched...may even find plasmids encoding antibodies from Addgene that you can make on your own.  To narrow your ... found this blog helpful be sure to check out Addgene’s blog for overviews of other antibody topics. ...10.4103/1947-2714.100998 Additional resources on the Addgene blog Find other tips and tricks for molecular ...

...which come from naturally-occurring immunoglobulin genes, recombinant antibodies are plasmid-based. Their... What is an Antibody? Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies...cascades that clear these infections. Anything that generates an antibody response in the immune system is referred...in the DNA of the hybridoma can occur, known as genetic drift, meaning that over time, the antibody you...monoclonal antibodies, recombinant antibodies are a homogeneous population. However, unlike monoclonal antibodies...antigen binding, the variable regions alone can be generated as a fusion protein. An ScFv is made up of the...detect the presence of a single protein from a heterogeneous mixture - for example, cell lysate or media ...

... 18 lung cancer patients. This study used CRISPR gene editing to knockout PD-1 on immune cells harvested... Cell heterogeneity Tumors are composed of a wide variety of cell types. This cell heterogeneity is an...cells that escape the elimination stage acquire genetic alterations and alter cell-surface antigen production... promotes the production of VEGF to increase angiogenesis around the tumor. This in turn regulates the... immunotherapy. Using activated DCs ex vivo to generate cytotoxic effector T cells, and targeting specific...played a huge role. For instance, researchers are genetically altering tumor cells and immune cells to knockout...element of successful tumor progression. A tumor’s genetic composition is constantly in flux and is dependent...

...proteins or cloning backbones to study your gene of interest with these tools. Conclusion Researchers and... Ligands GCE4All: Making Genetic Code Expansion Accessible Addgene's Genetic Code Expansion Collection... & Small Molecule Fluorophores. Addgene Blog. https://blog.addgene.org/better-dyeing-through-chemistry-small-molecule-fluorophores... is now the most popular self-labeling tag in Addgene’s collection. CLIP-tag: Also in 2008, Kai Johnsson...specific with a small and minimally-disruptive genetic tag on the protein of interest. Table 1: Summary...are the most popular self-labeling enzymes in Addgene’s collection, so you may be able to get a plasmid...ligand to use for your experiment, and explore Addgene's collection to find self-labeling fusion proteins...

...procedure and application to the major ALS/FTD disease gene C9ORF72. eLife, 8, e48363. https://doi.org/10.7554...101: Validation Resources on addgene.org Addgene's Antibody Guide Addgene's Antibody Protocols... outlined below. Knockout cell line approach Genetic knockout-based validation can conclusively prove...you are working with cells that are amenable to genetic knockout, this represents a very robust approach...to start with a screening procedure in easily genetically modified cell types (top). The specific approach...10.1038/s41598-019-55133-7 More resources from the Addgene blog Introduction to Gating in Flow Cytometry Antibodies...

...transition from weak, random binding to precision gene editing requires three major steps: RNA loading,...resources on addgene.org  Addgene's CRISPR Guide Addgene's CRISPR 101 eBook  Resources on the Addgene blog Components...from Streptococcus pyogenes.   Figure 2:  Crystal structure of S. pyogenes Cas9 in the apo state...    Figure 3: Crystal structure of S. pyogenes Cas9 bound to guide RNA (PDB ID 4ZT0), shown opaque...sequence isn’t in place yet.     Figure 4: S. pyogenes Cas9 in the apo state (PDB ID 4CMP, top) compared... residues in the CTD can get cozy with DNA. S. pyogenes Cas9 recognizes the PAM sequence 5’-NGG-3’, where...complementary strand is mismatched.   Figure 5: S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID...

...of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation." Nature biotechnology (2014). PubMed...to select a tool based on your project and not general opinion. I could spend a lot of time running through... Online tools dominate the software space but generally rely on less powerful algorithms to detect off-targets...tools available mostly work on any computer but generally require working knowledge of the command line ...tools require some kind of data input in order to generate results. While different types of input are more...focus on specific organisms. For this reason I generally recommend that laboratories that have already ...PubMed Central PMCID: PMC4262738. Resources at Addgene Get Tips on How to Design a gRNA Read Our CRISPR...

...). Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases...Over 75,000 pathogenic genetic variants have been identified in humans and cataloged in the ClinVar database...editing tools. We’ll summarize the major ones in Addgene’s repository later in the post. Advantages of prime...by double strand breaks has been widely used to generate precise changes. However, the efficiency of Cas9...Prime editing has a lot of promise for treating genetic diseases. In our summer Hot Plasmids post, we covered...demonstration of prime editing used to rescue a genetic disease phenotype in an animal model, which was...s41587-022-01527-4 Additional resources on the Addgene blog PRIDICT: Predicting Efficiencies of Prime...

...content on subjects such as: CRISPR Chemogenetics Optogenetics Genetic Code Expansion...Overview of Addgene's educational resources, including eBooks, science guides, videos, and protocols ... content , including protocols, tips on using Addgene’s materials, and career advice. Playlists include...Protocols Careers How-to Blog Read about plasmids in Addgene’s repository , and related molecular biology techniques...Concentrations for Bacterial Selection What is a Plasmid? Genetic Code Strain Information Molecular Cloning Techniques...

...excised, and gene expression proceeds. Gene Knockout: Placing the loxP sites on either side of a gene (called...shRNA expression. Gene Switch: These constructs contain two genes of interest, Genes A and B. When Cre... few popular recombinase-dependent gene expression strategies. Gene Expression: A stop codon flanked with...LSL" cassette) placed upstream of a gene of interest will prevent gene expression in the absence of Cre.... loxP"), will permit gene expression until Cre is present, at which time the gene will be disrupted or...Cre is absent, only Gene A is translated correctly. Cre expression excises Gene A and alters the reading...backbone to regulate your gene of interest. Depending on the construct, gene expression may be activated...

...Additional Gene Silencing Resources A modular plasmid assembly kit for multigene expression, gene silencing...plant-optimized plasmids for cloning, gene expression, reporter assays, genome editing, gene silencing, and more. Plasmid... a new window) — Tools for gene and protein expression and mapping of gene expression in plants. Grassius...Resources Cloning & SynBio Genome Editing Gene Silencing Reporters Addgene’s repository contains many plasmid ...from genome engineering and gene expression to reporter systems. Addgene’s collection includes plasmids...plasmids containing promoters that drive general and species-specific gene expression. Cloning and Synthetic ...most popular plasmids for gene editing in plants. Please refer to Addgene's Plant CRISPR Resources for...

...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Return to top Base Edit ID Plasmid Gene/Insert...Plasmid Gene/Insert Selectable Marker PI Publication Return to top Interfere ID Plasmid Gene/Insert Promoter...Plasmid Gene/Insert Promoter Selectable Marker Publication Return to top Prime Edit ID Plasmid Gene/Insert...Plasmid Gene/Insert Selectable Marker PI Publication Return to top RNA Knockdown ID Plasmid Gene/Insert...ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Return to top Epigenetics ID Plasmid ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Return to top CRISPR Resources Addgene has ...Marker PI Publication Return to top Purify ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication...

... Depositor Collections Gene Vector Core Viral Vectors Gene Vector Core Viral...remove genes flanked by either loxP or FRT sites respectively, and empty vectors to express your gene of ...and puromycin resistance gene from the CMV promoter pCDH-CMV 72265 Express gene of interest from the CMV...pCDH-EF1 72266 Express gene of interest from the EF-1 promoter pCDH-CB 72267 Express gene of interest from ...pCDH-CB-IRES-copGFP-T2A-Puro 72299 Express gene of interest from the CB promoter pCDH-EF1s 72484 Express gene of interest from ...recombinases. Genes can be cloned into these transfer vectors using shuttle vectors pICPIS-EF1 (Addgene #73355...College of Medicine Gene Vector Core Depositor...