We narrowed to 355 results for: Cre expression vector

...drive gRNA expression is dependent upon the cell type in question. In plant cells, gRNA expression is achieved...Cas9 expression is commonly driven by plant-derived RNA pol II promoters which regulate expression of longer...mRNAs for gene expression). Examples of commonly used RNA pol II promoters for Cas9 expression include the...carries several useful CRISPR toolkits for creating plant expression plasmids, including plant CRISPR plasmids...system in plant cells. Like other model systems, expression of S. pyogenes Cas9 or Cas9 variants (hereafter...OsU6 or OsU3, which are commonly used to drive expression of small RNAs in their respective species. Addgene...with other model systems, multiple gRNAs can be expressed to modify several genomic loci at once (get more...

...is_json="true" %}{% raw %}"Browse Empty gRNA Expression Vectors"{% endraw %}{% end_module_attribute %}{% ...href":"https://www.addgene.org/crispr/empty-grna-vectors/","type":"EXTERNAL"},"open_in_new_tab":true,"no_follow..., a specific base edit, or modulation of gene expression. Location and sequence are important considerations...CRISPRa and CRISPRi Finally, for modulating gene expression at the level of transcription – CRISPRa (activation...engineer specific DNA edits and to perform functional screens to identify genes involved in a phenotype of interest...sequence preferences must largely be ignored. Created with BioRender.com. The hammer: Gene knockout...terminus of the protein, to maximize the chances of creating a non-functional allele. For a 1 kilobase gene...

...plasmid contains humanized Cas9 expression cassette and sgRNA expression cassette. The target sgRNA can...a) Scheme of validation for DSB mediated EGFP expression cassette reconstitution. When the target sequence...annealing) took place and reconstituted the EGFP expression cassette. (b) pCAG-EGxxFP target plasmid and ... PX330 plasmids (or similar plasmids for gRNA expression), one gRNA at a time. After 48 hours of incubation...homology directed repair, and resulting in the expression of EGFP. Using this technique, you can assess...clone the PCR products into a pBluescript cloning vector and sequence the resulting plasmid. It can take...mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA." Scientific reports...

...tracrRNA was combined with the sgRNA expression vector allowing the screen to be conducted with only two constructs...gain of function screens. These screens can help reveal the function of lowly expressed genes or genes ...regulate gene expression. Some CRISPR libraries contain many sgRNAs per gene to increase confidence in... sgRNA library In addition to inhibiting gene expression, dCas9 can also be used for transcriptional activation...protein-tagging system for signal amplification in gene expression and fluorescence imaging." Cell 159.3 (2014):...Calabrese library to screens performed with open reading frame (ORF) overexpression libraries for resistance...large-scale functional screens Read this blog post about genome-wide screening using CRISPR Resources...

.... They started by delivering an AAV construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive...potential afterwards. This measurement trended with expression of the opsin in the target neurons, validating...studies Channel rhodopsins for optogenetics Viral Vectors 101: Chemogenetics ... intestines known as neuropods. They observed increased vagus nerve firing in response to both of these...

... al., 2016). Using lentiviral vectors from Addgene, the lab will express these candidate genes in TS iPSCs... It’s possible that solute channels typically expressed on other cell types can compensate for or bypass...defects. Eight genes from the X chromosome are expressed at significantly reduced levels in women with ... Association for Creatine Deficiencies: Compensating for SLC6A8 mutations in Creatine Transporter Deficiency...SLC6A8 gene, also known as the creatine transporter. The Association for Creatine Deficiencies (ACD) is a parent-let...regeneration of ATP from ADP is creatine. In order to reach the brain, creatine needs to be transported from...Deficiency Creatine Transporter Deficiency (CTD) is a rare X-linked inherited neurometabolic condition and...

...Conditional (Cre-lox), stable expression of shRNAs; addition of Cre turns on shRNA expression. See Ventura...long-term transgene expression. Wild-type lentiviruses have been engineered into viral vectors for safe use ...Lentiviral Vector Guide Popular Lentiviral Transfer Plasmids Looking for lentiviral plasmids to express a CRISPR...cDNA expression. David Sabatini 14883 FUGW 3rd hUbC-driven EGFP. Can be used for cDNA expression. David... constitutive transgene expression. Contains intron that gives high expression. Didier Trono 12254 pWPI...plasmid for cDNA expression. Miguel Ramalho-Santos 24129 pULTRA 3rd Bicistronic expression of EGFP and the...pHIV-EGFP 3rd EF-1alpha driven expression of cDNA and and EGFP co-expression. See plasmid 21374 for dTomato...

...Cells You may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference...transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene of interest Triton X-100 Benzonase/...harvesting. The addition of sorbitol has been shown to increase viral titers by up to 1.8-fold ( Rego et al., ...sodium bicarbonate, sorbitol has been found to increase viral titers by up to 1.8-fold. See the recipe...

...in therapeutics, as it can increase gene expression at certain loci to create changes in phenotypes. For...activate, we would recommend using a cDNA overexpression vector rather than going through all the troubleshooting...: once the environmental trigger leaves, gene expression is no longer activated. However, there are several...up-regulated. For instance, generally, the more highly expressed a gene is under native conditions, the less activation...libraries, which consist of plasmids that over-express coding sequences from a given cell type or organism... for altering DNA sequence or regulating gene expression.     References Behan FM, Iorio F, Picco G, Gonçalves...Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging. Cell 159:635–646 . ...

...subset of RSMs that regulate gene expression) that uniquely express selectable and/or sortable reporters...programs that specify which genes should be expressed or not expressed in each state. To help researchers adapt...register”. Each chemical input to an RSM drives expression of a distinct large serine recombinase that recognizes...system), these reactions are irreversible. Once expression of a particular recombinase is triggered, its...possible downstream states that can be achieved by expression of other recombinases. Multiple orthogonal attB-attP...present a two-input, five-state GRSM register that expresses a unique combination of red, blue, and green fluorescent...state detection with flow cytometry. However, expressing reporters will consume cellular resources and...

...loxP sites such that their expression is turned on or off in cells expressing Cre. The result of a system...15 years efforts have been made to create transgenic mice expressing fluorescent protein reporter constructs...allows the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell...immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes...above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after...useful tool for marking and tracking cells is the the Cre/lox recombination system. A researcher can flank ...respectively. The permanent labelling of cells using cre-lox is advantageous when compared to photoactivable...

...avoid accidentally creating a gene drive is also the easiest: don't use a DNA vector that encodes both ...spread exclusively through populations that already express Cas9 from an unlinked locus, while a Cas9+sgRNA...attempting to build gene drives in tropical mosquito vectors of diseases such as malaria and dengue are currently...Cas9 should be aware of the potential hazards of creating “gene drives” capable of spreading through wild...does making transgenic organisms with Cas9 risk creating a gene drive? The simplest way to make a gene ...appropriate flanking homology into a germline cell can create an RNA-guided gene drive. In principle, this event...

...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to ... closed RNAs seem to be conserved in mammals, expressed in neurons, and are especially abundant in neuronal...resulted in a cell migration defect. They have created a circRNA website so scientists can view public...developments. Using the Tet-on system, his lab has created a tetracycline cas9 inducible knock-out system....

...is then digested and ligated into an appropriate vector. Gibson Assembly, GeneArt Cloning, and SLIC can...of recombineering. In recombineering, bacteria express phage recombination machinery, which facilitates...products are directly transformed into bacteria expressing recombineering proteins Red/ET from a temperature-sensitive...harnesses the power of bacterial recombineering to create insertions, deletions, and substitutions - at the...

...constitutive expression as well as those that allow for conditional (Cre-lox) or inducible (Tet) expression are...shRNAs are often delivered using viral vectors. Lentiviral vectors are a preferred method for long-term ...Plasmid Vector Type PI Publication Additional Resources Addgene Resources pLKO.1 cloning vector protocol...interference. Created with BioRender.com. shRNA Plasmids Browse our selection of empty vectors for cloning...Addgene; browse by constitutive lentiviral shRNA, cre-regulated lentiviral shRNA, and more. Plasmid...long-term gene silencing. Once the viral vector is taken up by a host, the viral genome is transcribed, producing...Lentiviral packaging plasmids Web Resources Trono Lab Lentivectors Toolbox Optimized shRNA design in pSico with...

... promoter to drive expression of a fluorescent gene and packaged into viral vectors that can be administered...non-coding sequences of DNA that regulate gene expression by interacting with proteins such as transcription...enhancers are functional in every cell. Differential expression of transcription factors, which recognize and...mechanism that allows enhancers to regulate gene expression in a cell type-specific manner also makes them...tools for targeting those specific cell types to express exogenous genes — a feature the AIBS has capitalized...Genetic Tools Atlas. To evaluate the resulting expression of the fluorescent reporter, the brain tissue...Functional enhancer elements drive subclass-selective expression from mouse to primate neocortex. Cell Reports...

...stability. Additionally, the choice of vector can impact the expression level of an antibody (Ayyar et al....such as parasites or microbes, or to proteins expressed on the surface of cells that have been infected...the protein out of solution, or isolate cells expressing this protein. Read on to learn more about antibodies...context of the immune system, each isotype is expressed at different times of the immune response and ...qualitative and quantitative measurement of protein expression in cell lysates, whole cells, or tissue samples...frequently used to compare relative levels of protein expression between cell types or treatment conditions. They....    Figure 4: Western blot showing GFP expression in cells that have been transfected with a plasmid...

...the phage display screens are subcloned (using FX cloning) into the expression vector pSb-Init, which attaches...their small size, nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target...sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins...to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure 1: ...threonines at positions that can be randomized for creating the libraries (Zimmermann et al., 2018). Find ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...the candidate sybodies in their coat protein to screen for binding to the target protein. The protocol...

...consist of a group of genes/proteins that, when expressed, normally encourage cell survival, growth and ...Clone Collection 2.0. The collection is a set of vectors for use with the Gateway Cloning Platform (Life... is that it enables the simple construction of vectors to study the function of any of these genes through...as 30% of all cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations...he or she can use to study it (as seen in the screenshot below). Currently, the page includes interactive... Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer Library Can Be Used...

...flow cytometry, and mRNA expression.   If your protein of interest is expressed at similar levels in ...useful to produce a sample with mixed expression, with the expression measurable through an epitope tag such... sets, you can increase confidence that the labeling observed represents the expression level of the target...epitopes. Detection of overexpressed (tagged or untagged) protein When expression plasmids are available...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...caused by overexpression. Additionally, if the cell line used has endogenous levels of expression, this can...preferred. The overexpression approach is therefore best suited to cell lines without any expression of the target...

...These proteins can be expressed in specific neurons, often utilizing viral expression systems, to activate...Insert Vector Type Promoter PI Return to top Additional Resources Optogenetics Guide Viral Vector Packaging...identified to increase the spectral and kinetic properties as well as improve the expression and membrane...fluorescent protein to the opsin in order to track its expression and localization. Many other types of proteins...inhibition. For example, search for "ChR mCherry", "Cre inhibition", or other terms. Looking for viral preps...optogenetics plasmids. ID Plasmid Description Gene/Insert Vector Type Promoter Eligible for Viral Packaging PI Return...

...Viral Vector Packaging Service AAV Recombinases Viral Vector Packaging Service: Recombinases Cre AAV Dre...recombinases, which can be used to control gene expression. See our Cre-lox Plasmids page for more information ...Deisseroth 55636 pAAV-EF1a-Cre EF1a none 11, rg* Karl Deisseroth 121675 pAAV-EF1a-fDIO-Cre EF1a none 1, 5, 8, ...pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 CMV eGFP 1, 2, 5, 8, 9 James M. Wilson 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a mCherry... Cepko 69916 pAAV.cTNT.iCre cTnT tdTomato 9 William Pu 203842 pAAV-Ef1a-fDIO-eGFP-2A-Cre-WPRE EF1a EGFP...Spiegel 105550 pAAV.GFAP.Cre.WPRE.hGH GFAP none 5, PHPeB James M. Wilson 196410 AAV-GfaABC1D-Cre-4x6T GfaABC1D...Jonathan Ting 107788 AAV.rTH.PI.Cre.SV40 rTH none 9, rg* James M. Wilson 24593 AAV-pgk-Cre PGK none rg* Patrick...

...that the percentage of dystrophin-expressing muscle cells increases over time, and all three groups confirm...myoblasts; when injected into DMD mice, these cells expressed functional dystrophin. Editing genes in vivo is..., the CRISPR components would only need to be expressed for a short period of time, which would prevent...confirm dystrophin expression via Western blot, at <10% of the wild-type level. Muscle histology is improved...stem cells 2. Remove any immunogenicity of the AAV vector Once delivery has been optimized, it’s important... reduce off-target editing. Short-term CRISPR expression would be especially desirable, as it would reduce... genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy. Science...

...marvel of DNA compaction, an enhancer can increase the expression of a gene that is millions of base pairs...genome math expression for NANOG enhancers, we’re ready for step 2: design a CRISPR screen to disrupt ...CRISPR screen. We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an...Putting it all together, we get the following expression for step 1: (H3K4me1 + H3K27ac) – H3K4me3 The...stem cells, and conclude with a targeted CRISPR screen that can disrupt these enhancers. A real-world ...that have NANOG binding sites 2. Design a CRISPR screen to target and disrupt the NANOG enhancers 3. CRISPR...you’re interested in designing your own CRISPR screens for enhancers, check out the Appendix. Conclusion...

...cells expressing Cre AND Flp, here expressed through a double transgenic animal strategy (PV-2a-Cre; SOM-IRES-Flp...only Cre AND Flp, Cre NOT Flp, or Flp NOT Cre. B) This strategy has been expanded to incorporate VCre in...EYFP Cre AND Flp Yes 231926 pAAV-Ef1a-Con/Fon-EYFP Cre AND Flp No 55651 pAAV-hSyn Con/Foff EYFP Cre AND...AND NOT Cre Yes 137129 pAAV-Ef1a-Con/Fon-BFP Cre AND Flp Yes 137130 pAAV-Ef1a-Con/Foff 2.0-BFP Cre AND NOT.../Von-EYFP Cre AND Flp AND Vcre Yes 137165 pAAV-Ef1a-Con/Fon/Von-GCaMP6M Cre AND Flp AND Vcre No References...intersectional expression plasmids that combine multiplexed recombinases, including Cre, Flp, and VCre to enable...Intersectional expression approaches combine multiplexed recombinases, including Cre, Flp, and VCre to enable...

...deliver in low-capacity vectors, such as adeno-associated viral (AAV) vectors. In 2017, Zetsche et al....multiplexing approach using a single crRNA array to express up to 4 crRNAs. Zetsche et al. also suggest that...PAM site, Cpf1 cleaves in a staggered fashion, creating a 5 nucleotide 5’ overhang 18-23 bases away from...mediated editing, as Kleinstiver et al. saw no decrease in Cpf1 activity with 4-6 base deletions at the...

... empty backbones, fluorescent proteins, and Cre expression plasmids. Large collections or kits are described...General Tools Fire Lab C. elegans Vector Kit - Andrew Fire Lab. A set of vectors for C. elegans research, including...and spatial control of gene expression in C. elegans by combining expression of the FLP recombinase from...can knock down genes by feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily...developed and characterized which allow researchers to express desired constructs in precise cell types and/or...ID Plasmid Description Gene/Insert PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids...plasmids intended for use together (e.g. cloning and expression systems) and therefore supplied as one item. ...

...al., 2024). In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study...than SpyCas9, allowing them to be packaged in AAV vectors, potentially with multiplexed targets.   ...showed that AAV-delivered CHARM could silence the expression of prion protein in mouse brains for at least...gene product; it simply turns the target protein expression off. Plus, DNA methylation is a long-lasting ...-silencing CHARM variants that stop their own expression after silencing their target — potentially improving...by targeting marker proteins that are uniquely expressed in a given cell type. For example, glial fibrillary...GFAP) is a glia-specific intermediate filament expressed in astrocytes throughout the brain, making it ...

... lentiviral packaging systems, and luciferase expression. On individual material pages, we added curation...curation fields for purpose, promoters, expression types, and more, all of which help a researcher understand... Inducible Plasmids Empty Backbones Bacterial Expression Systems AAV Plasmids Blogs Origin of ... CRISPRTwenty Years of Sharing: Addgene's Viral Vector Service...ve learned that increased accessibility is achieved through interconnected incremental changes. It was...When Addgene opened in 2004, we focused on creating a repository that made it easy to share plasmids,...identified and offered a number of solutions that increased accessibility, such as using couriers that refill...

...the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...for quantitative imaging, Ming Hammond’s group created Spinach2 using site-directed mutagenesis of Spinach...for in vivo studies. Dimerization of Broccoli increases its fluorescence by about 1.8-fold, and may be...for greater contact with its RNA aptamer. This increased interaction between Corn and its small molecule... to TO1-biotin. Two of these variants exhibit increased brightness compared to Mango, and all are photostable...

... groups was to create a system that would use light to turn on and off gene expression while imparting...HEK293T cells to induce expression of human IL1RN, the researchers saw an 11-fold increase in mRNA levels after...Phillips-Cremins JE (2019) LADL: light-activated dynamic looping for endogenous gene expression control...transcriptional activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the...exposure (470nm), the researchers showed that expression of the human ASCL1 gene could be spatially controlled...demonstrated the ability to spatially control gene expression. Both of these techniques take advantage of light-induced...light-activated split-Cas9 (FAST) system constitutively expresses the C-terminal fragment of Cas9 while transcription...

...targeting vector assembly, and the remaining 5 plasmids include Cre and FLP expression vectors, a general... one vector from the Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_T2 (Addgene #79888) , which expresses an untagged...gene-specific CRISPR and donor vectors as well as pX330S-2-PITCh, expressing a generic gRNA targeting the...Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA, & FBL-specific...are available. An improved FLP recombinase expression vector allows for efficient marker cassette FLP-out...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors By Species Mammalian Bacteria...has been adapted to allow tagging of proteins expressed from their natural chromosomal context. These ...

...changes in root-to-shoot ratio in Arabidopsis, (2) expression patterns in plants with enhanced sodium accumulation...protocols Find eBooks on plasmids, CRISPR, viral vectors, and more ...the community, increasing the impact of the work, and as a collateral benefit, increasing the impact of...Tompson Institute.  References Baldwin, Melinda. "Credibility, peer review, and Nature, 1945–1990." Notes and...

...Microbiology Plant Expression Stem Cells Synthetic Biology Tet Inducible Expression Worm Expression Kits Kits ... Fluorescent Proteins Protein Expression Synthetic Biology Viral Vectors Communities Find a scientific...Genome Engineering Genome Engineering Overview CRISPR Cre-lox TALENs Zinc Fingers Luminescence Fluorescent ... Topics Antibody Plasmid Collection Bacterial Expression Systems Empty Backbones Genetic Code Expansion...libraries. Pooled Library Types Barcoding CRISPR Screening Surface Display cDNA Depositor Collections A modular...

...cells Peter Cockerill 212936 pGL3 Basic Vector Firefly Vector for investigating regions controlling transcription...a reference level of expression. Browse or search for popular plasmids expressing luciferase. We also ...Mammalian expression of Nano-lantern Takeharu Nagai 87121 pcDNA-RLuc8 Renilla CMV Mammalian expression Renilla...CMV AAV expression of firefly luciferase and GFP James Wilson 22522 phGluc Gaussia EF1α Expression of Gaussia...Firefly TRE Lentiviral vector with dox- or tet-inducible luciferase expression Stephen Tapscott 83281 ...Mammalian expression of firefly luciferase Lee Helman 131860 pZmUBQ:LUC Firefly ZmUBQ Plant expression of firefly...Renilla CAG AAV expression of Renilla luciferase Mark Kay 83280 pscAAV-CAG-RLuc Renilla CAG Vector for packaging...

...pitfalls: they require construction of plasmid vectors and/or rely on cumbersome in vitro synthesis of...purchased and there is no need to construct plasmid vectors or synthesize/purify IVT sgRNA in the lab. Figure... (although we could successfully knock-in non-expressed genes with similar efficiency, Dewari et al. 2018...tagging success, although many of these genes were expressed at really low levels so actual knock-in success...antibody” can be used for downstream IP/ChIP assays. Creating epitope-tagged clonal cell lines remains a challenge...crRNA complementary to the template strand could increase knock-in efficiency.Note: Our TAG-IN tool can ...

...targeted repression of gene expression. They successfully knock down expression of essential and non essential...) technology (Ronda et al., 2016). pMA7CR_2.0 expresses lambda Red and Cas9, which are separately inducible...function in bacteria, most efforts to regulate gene expression were limited to inducible promoters or direct... much more user-friendly way to modulate gene expression. The Marraffini lab and Qi lab developed early...repression. Using a Tet responsive promoter to drive expression of a catalytically dead Cas9 enzyme allows reversible...the gRNA cassette, one can reliably knockdown expression of multiple target transcripts simultaneously...genes in M. tuberculosis. dCas9 and a gRNA are expressed on pRH2502 and pRH2521, respectively. Both dCas9...

... Flp-dependent Cre and Flp-dependent Cre, Flp, and VCre-dependent Constitutive (non-cre-dependent) Clear... Viral Vector Packaging Service AAV Controls Viral Vector Packaging Service: Control...None Other Cre-dependent color switch Brainbow constructs Spaghetti monster tags Activity Cre-dependent...floxed-mCherry-WPRE-HGHpA EF1a mCherry Cre dependent 9 Karl Deisseroth 27056 pAAV-Ef1a-DIO EYFP EF1a EYFP Cre dependent 1, 2,...EF1a mScarlet Cre dependent 1, 5 Karl Deisseroth 50457 pAAV-hSyn-DIO-EGFP hSyn EGFP Cre dependent 1, 2...mCherry Cre dependent 1, 2, 5, 8, 9, rg* Bryan Roth 51502 AAV pCAG-FLEX-EGFP-WPRE CAG EGFP Cre dependent...pAAV-CAG-FLEX-EGFP CAG EGFP Cre dependent 1 Ian Wickersham 104052 pAAV-CAG-DIO-EYFP CAG EYFP Cre dependent PHP.V1...

...search scoped to plasmids shows filters for expression and vector type, while a search scoped to viral preps...well-characterized. During this time, we’ve also created a variety of educational resources in an effort...understanding of Addgene’s customer’s needs. Categories We created a categorization of our information in order to...for promoter and serotype, among others. We also created a normalization mechanism to “collapse” variations...

...Intersectional viral expression of GCaMP6m/s in cells depdent on Cre, Flp, or Vcre Comprehensive Dual- ... Calcium Intersectional viral expression of sRGECO (constitutive or Cre- or Flp-dependent) Comprehensive...Adenosine Express the adenosine sensor GRAB_Ado1.0 in neurons or astrocytes (Constitutive or Cre-dependent...cyclic AMP (cAMPr) for mammalian or insect expression (Cre-dependent) cAMPr: A single-wavelength fluorescent...iSeroSnFR for bacterial or mammalian expression (Constitutive or Cre-dependent) Directed Evolution of a ...Genetically Encoded Calcium Indicators (Constitutive or Cre-dependent) jGCaMP8 Fast Genetically Encoded Calcium...High-performance GFP-based calcium indicators (Constitutive or Cre-dependent) High-performance calcium sensors for imaging...

... Plasmid Collections Bacterial Expression Systems Bacterial Expression Systems Protein Purification Protein...bacterial expression systems. Read below and browse our curated collection of bacterial expression plasmids...Addgene Blog E. Coli Strains for Protein Expression Protein Expression Epitope Tags Protein Tags Return to ...to top Controlled Expression It isn’t always advantageous to constitutively express your gene of interest...turn on expression only at a specific time. This collection contains plasmids whose expression levels ...Addgene Controlled Expression Resources Check out our Tetracycline (Tet) Inducible Expression Collection for...Find bacterial expression plasmids that can be used for protein purification, protein visualization, ...

...Gurumurty et al., 2016). AAV can also be used as a vector to deliver CRISPR/Cas9 either through injection...Mediated Genome Editing with Adeno-Associated Viral Vector. iScience 9:286–297 . https://doi.org/10.1016/j.isci...Resources on the Addgene Blog CRISPR 101: Mammalian Expression Systems and Delivery Methods Learn about Mouse...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...knock in models). CRISPR mouse model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide...selection offered by mouse ESCs. For example, we often screen only 15-25 mice when generating knockout models...for generating mouse models Although CRISPR is incredibly useful for generating mutations by NHEJ and generating...

...drawback of CRISPR-Cas9 technology is that most expression systems are constantly “on”. There is no mechanism....org  View Addgene's collection of empty gRNA vectors View Addgene's CRISPR resources Request ready-to-use... or phosphate molecules of the RNA backbone to increase stability.  Nature has evolved ways to prevent...surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency. In the same vein, locked...

...Tetracycline (Tet) transactivator to increase fluorescent protein expression C234-E25: attB2r-SNAP-attB3 SNAP-tag...can also assemble vaccines, target gene therapy vectors, or lock together smart biomaterials. There are...assembly tools, alongside criteria for Click Biology. Created with BioRender.com.   Principles of Click Biology...Protein (MFP) through rapid covalent coupling. Created with BioRender.com.   Table 1: A handful of...Cas9 plasmids with N- or C-terminal PCV2 HUH-tag; increase homology-directed repair efficiency by linking...linking the repair template VirD2 HUH-tag For creating DNA-protein conjugates rHUH HUH-tag (RNA) Covalent...

...viral vectors (here and here).  Figure 3: JEDI-2P was identified from a high-throughput screen, selected...Figure 2: iGluSnFR3 displays enhanced brightness, expression, and sensitivity compared with earlier SF-iGluSnFR.... By fusing ITAM to an RTK of interest and co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence... They use a unique and powerful high-throughput screen to select variants based on performance metrics...resulting JEDI-2P has faster response kinetics and increased brightness, sensitivity, and photostability under...selected for increased voltage response, brightness, and photostability. It enables sustained deep-tissue... be generalized to other RTKs of interest, they create sensors for FGFR1, EGFR and ErbB2. And orthogonal...

...cloning vector Retroviral vectors: neomycin (pSIR-neo) , GFP (pSIR-GFP) , DsRed (pSIR-DsRed-Express2) , human... sgRNA dual expression: pAC2 , pAC152 , pAC153 , pAC154 ; pmax dCas9 activator expression: pAC91 , pAC92... pooled library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro...Addgene Church gRNA design and cloning gRNA cloning vector PDF, 118 KB Church gRNA design and cloning for ...pCFJ90 (pharyngeal co-injection marker); pDD104 (Cre recombinase) PDF, 105 KB Jaenisch CRISPR plasmid ...human pluripotent stem cells pCas9_GFP ; gRNA empty vector Protocol at StemBook O'Connor-Giles Fly: gRNA and...PDF, 330 KB Parrott NEBuilder Assembly of CRISPR vectors using ssDNA oligos p201G Cas9 ; p201B Cas9 ; p201H...

...UTRs are included in the vector to enhance expression of the message. Both vectors utilize homodimeric FokI...contains modified pFUS array vectors, destination vectors and a reporter vector for mammalian cell-based ...Destination vectors, pcDNA-TAL-NC2 and pCAGGS-TAL-NC2, are mammalian expression/mRNA synthesis vectors containing...much higher than the parental vectors. pC-GoldyTALEN directs expression of TALENs from a truncated CAGs...placing TALORs built into this vector under galactose-inducible expression. pTAL6-BB contains the TEF1 promoter...promoter, giving constitutive expression of TALORs built into this vector. 36034 pTAL6-BB Golden Gate TALEN...1214–2210 of pTAL3 were cloned into a pCS2 expression vector resulting in shorter N- and C-terminal tal...

... vitro, and can be used for measuring protein expression or localization in cells, or even single-particle...Mol Syst Biol. 2014 Galit Lahav EYFP Gateway vectors with EYFP split at 175 Arabidopsis homolog of the...acetylation system and novel in vivo high-throughput screen for molecules affecting alpha-synuclein oligomerisation...

...species-specific gene expression. Cloning and Synthetic Biology Expression Vectors The table below highlights... and collections are designed for creating plant expression vectors. ID Kit Description Cloning Type PI...Resources Improved Gateway binary vectors — High-performance vectors for creation of fusion constructs in transgenic...MultiGreen Kit Modular assembly of multi-gene expression vectors. Golden Gate (GreenGate) Wayne Parrott Additional...maize and Arabidopsis) and vectors for easy assembly of one or more gRNA expression cassettes. A CRISPR/Cas...multigene expression, gene silencing, and silencing rescue in plants. JoinTRV , an improved vector system... — Tools for gene and protein expression and mapping of gene expression in plants. Grassius (Link opens...

...Mammalian Expression HcRed1-N1 - Mammalian Expression HcRed1-C1 - Mammalian Expression dKatushka 588 635 22...pBad-EBFP2 - Bacterial Expression EBFP2-N1 - Mammalian Expression EBFP2-C1 - Mammalian Expression EBFP2-pBAD - ...- Mammalian Expression pCyPet-C1 - Mammalian Expression pCyPet-His - Bacterial Expression CYPet-N1 - Mammalian...MiCy-N1 - Mammalian Expression MiCy-C1 - Mammalian Expression MiCy-pBAD - Bacterial Expression Return to top... - Mammalian Expression sfGFP-C1 - Mammalian Expression sfGFP-pBAD - Bacterial Expression moxGFP 486 510...Mammalian Expression pCAG-GFP - Mammalian Expression pLV-eGFP - Mammalian Lentiviral Expression Ac5-STABLE1...1 - Mammalian Expression mEGFP-N1 - Mammalian Expression mEGFP-C1 - Mammalian Expression mEGFP-pBAD - ...