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We narrowed to 370 results for: Cre expression vector

Showing: 241 - 280 of 370 results

  1. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ... 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...

  2. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...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 . ...

  3. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...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...

  4. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...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...

  5. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...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...

  6. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...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....

  7. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...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...

  8. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    ... 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...

  9. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...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...

  10. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...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...

  11. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...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...

  12. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...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...

  13. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...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...

  14. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...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...

  15. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...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...

  16. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...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 ...

  17. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ... 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...

  18. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...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...

  19. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ... 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...

  20. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ... plasmids expressing Cre recombinase and filter the results by expression system and vector type. Find...or control gene expression or skip ahead to browse highlighted plasmids expressing Cre , Flp , Dre , other...Summary of Cre, Dre, and Flp Recognition Sites. Controlling Recombinase Expression By expressing the enzyme...interest will prevent gene expression in the absence of Cre. In the presence of Cre, the stop codon is excised...lox-STOP-lox shRNA constructs, Cre expression promotes shRNA expression. Gene Switch: These constructs...Genes A and B. When Cre is absent, only Gene A is translated correctly. Cre expression excises Gene A and...double-crossover event. Toxicity Cre-lox recombination is a very useful tool, but Cre expression can be toxic in some...

  21. Brain Initiative Collection

    Type
    Collection
    ...CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA AAV vector for Cre expression in forebrain GABAergic interneurons...-AAV5 pAAV-CAG-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...AAV8 pAAV-Ef1a-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...AAV9 pAAV-Ef1a-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...combination with Cre-dependent transgenic mouse lines or co-infection with Cre dependent viral vectors. Jonathan...AAV-mediated expression of voltage sensor under the Syn promoter, Cre-dependent expression; soma localization...

  22. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    ...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...

  23. Zhang Lab CRISPR Page

    Type
    Collection
    ...2015). The dual vector system uses S. pyogenes Cas9 (SpCas9), using one vector to express SpCas9, and another...delivering Cre and sgRNA to a Cre-dependent mouse or sgRNA to a constitutively Cas9-expressing mouse. Described...luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone with luciferase-2A-Cre recombinase Vectors for brain...mouse toolbox protocol 637.8 KB CRISPR-Cas9 Cre expression vectors for cancer modeling Using Cas9 mice, Platt...pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR This plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase...target site sequence (20bp). CRISPR-Cas9 Cre expression vectors for genome editing in the brain Using Cas9...

  24. Kazuhiro Oka Lentiviral Vectors

    Type
    Collection
    ...Collections Gene Vector Core Viral Vectors Gene Vector Core Viral Vectors Lentiviral Transfer Vectors Adenoviral... transfer vectors and generating new viral vectors. Download the Lentivirus Transfer Vector Production...Lentiviral Vector ID Purpose pCDH-EF1-DIO-copGFP 72253 Expresses copGFP under EF-1 promoter when Cre is expressed...by another vector pCDH-EF1-DIO-tdTomato 72254 Expresses tdTomato under EF-1 promoter when Cre is expressed...promoter when FLP is expressed by another vector pCDH-EF1-Fon-tdTomato 72261 Expresses tdTomato from the ... promoter when FLP is expressed by another vector pCDH-EF1-FLPe 72262 Expresses FLPe from the EF-1 promoter...lentiviral and adenoviral vectors read our Lentiviral Vector Guide and Adenoviral Vector Guide Find more plasmids...

  25. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ...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 ...

  26. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...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...

  27. AAV Molecular Tools

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    Collection
    ...FLEX-tdTomato-T2A-SypEGFP-WPRE Syn-driven, Cre-dependent Cre-dependent expression of cytoplasmic tdTomato and synaptophysin-EGFP...2A-Synaptophysin-mRuby Syn-driven, Cre-dependent Cre-dependent expression of membrane-bound GFP and synaptophysin-mRuby...Name Expression System Activity Serotype PI 124364 pAAV-FLEX-DTR-GFP CBA-driven, Cre-dependent Cre-dependent...pAAV-flex-taCasp3-TEVp EF1a-driven, Cre-dependent Cre-dependent, bicistronic expression of designer pro-taCasp3 and...pAAV-FLEx-ER-TurboID CMV-driven, Cre-dependent Cre-dependent expression of ER-localized TurboID (with HA...-mKate2-PDE4D3-Cat EF1a-driven, Cre-dependent Cre-dependent expression of mKate2 and a truncated human...pAAV-FLEX-P301L Tau EF1a-driven, Cre-dependent Cre-dependent expression of human 2N4R microtubule-associated...

  28. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    ...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...

  29. Retrograde AAV viral preps

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    Collection
    ...Color-flipping switch. Expresses NLS-mCherry in the absence of Cre, and expresses NLS-EGFP in Cre-positive cells...AAV-pgk-Cre PGK Cre expression Recombinases Patrick Aebischer 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a Cre...physically-separate mCherry expression Recombinases Karl Deisseroth 55636 pAAV-EF1a-Cre EF1a Cre expression Recombinases...-tagged Cre expression Recombinases James M. Wilson 105553 pENN.AAV.hSyn.Cre.WPRE.hGH Syn Cre expression...pENN.AAV.CamKII 0.4.Cre.SV40 CamKII Cre expression Recombinases James M. Wilson 107738 pAAV-hSyn-Cre-P2A-dTomato...pAAV-hSyn-Cre-P2A-dTomato Syn Cre expression with simultaneous dTomato expression Recombinases Rylan Larsen 107788 AAV.rTH.PI.Cre.SV40...EF1a Flpo expression Recombinases Karl Deisseroth 55638 pAAV-EF1a-vCre EF1a VCre expression Recombinases...

  30. Tetracycline Inducible Expression

    Type
    Collection
    ...85040 pK170.AAV-TRE-Cre-WPRE (Supernova) AAV vector for dox-inducible expression of Cre recombinase. Takuji...63704 pRetroX GFP T2A Cre Retroviral vector for dox-inducible expression of GFP T2A Cre recombinase. Floris...Neo-TRE-CMV-Cre-rtTA Dox-inducible expression of Cre recombinase. Madeline Lancaster 198752 DiLiCre 2.0 Dox-inducible...popular empty backbones, expression vectors for transactivators, and vectors for other applications, or...Lentiviral Tet-Off vector for repressible expression with blasticidin selection. *Note: this vector contains an...PB-EF1a-TetOn3G PiggyBac vector expressing Tet-On 3G transactivator constitutively expressed by EF1a promoter ...Tetracycline (Tet) Inducible Expression Tetracycline (Tet) Inducible Expression You may also like... Blog...

  31. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...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...

  32. Caltech Systemic Capsids

    Type
    Collection
    ...Cre-expression Cre Wilson 107738 pAAV-hSyn-Cre-P2A-dTomato Syn Cre-expression Cre Larsen 140135 pAAV-EF1a-iCreV...pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 Syn Cre-EGFP expression Cre Wilson 105550 pAAV.GFAP.Cre.WPRE.hGH GFAP Cre-expression... Viral Vector Packaging Service AAV Caltech Systemic Capsids Viral Vector Packaging Service... a new window) . Viral Vectors 101: Systemic Capsids PHP.eB These viral vector preparations were produced...Activator, Cre-dependent DREADD Roth 44362 pAAV-hSyn-DIO-hM4D(Gi)-mCherry Syn Inhibitor, Cre-dependent ...sensor, Cre-dependent GCaMP Kim , GENIE 104492 pGP-AAV-syn-FLEX-jGCaMP7f-WPRE Syn Calcium sensor, Cre-dependent...sensor, Cre-dependent GCaMP Kim , GENIE 104496 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE CAG Calcium sensor, Cre-dependent...

  33. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...compatible Y2H vector - BD pRPR1_gRNA_handle_RPR1t - gRNA expression empty vector pFA6a vectors - Epitope ...lentiviral vector for ORF expression pDest-565 - Gateway destination vector for bacterial expression; His,GST...Empty gRNA Expression Vectors page Cre-lox Site-specific recombination See our dedicated Cre-lox Plasmids...gene expression vector URA3 Yeast pAG306GAL-ccdB - Yeast Gateway expression vector (see...Inducible Expression page Bacteria Lac, T7, araBAD, trp mTagBFP2-pBAD - Protein expression vector with C-...versions of pFastBac LIC vectors and pFastBac Dual LIC vectors for insect expression Fluorescent proteins ...Bacterial expression of EBFP2 blue fluorescent protein 35S-eGFP-nosT - Transient expression vector for eGFP...

  34. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...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...

  35. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...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...

  36. Brain Armamentarium

    Type
    Collection
    ...CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA AAV vector for Cre expression in forebrain GABAergic interneurons...combination with Cre-dependent transgenic mouse lines or co-infection with Cre dependent viral vectors. Jonathan...pAAV-AiE0452h-minBG-iCre(R297T)-BGHpA (Alias: CN3781) Enhancer AAV for Cre recombinase expression in striatal ..._3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN3779) Enhancer AAV for Cre recombinase expression in striatal ...pAAV-AiE0450h-minBG-iCre(R297T)-BGHpA (Alias: CN3778) Enhancer AAV for Cre recombinase expression in striatal ...version of DLX enhancer). AAV vector for strong & rapid SYFP2 expression in forebrain GABAergic interneurons...pAAV-AiE0441h_3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN4825) Expression of iCre(R297T) in striatal medium...

  37. Zebrafish Plasmid Collection

    Type
    Collection
    ...fluorescent proteins, reporters, biosensors, and Cre expression plasmids. Large collections or kits are described.... pGCS Vector Kit - Hai-Ning Du Lab. A set of pCS2+ backbone-based Gateway destination vectors (pGCS),...which an exogenous mRNA can be injected and expressed. EXPRESs Plasmids for Zebrafish Receptors and Zebrafish...in zebrafish. Zebrafish expression atlas - A zebrafish-specific gene expression database that provides ...resistant cell types. CreLite - George Eisenhoffer Lab. An optogenetically controlled Cre/loxP system that ...A useful “starter kit” for screening fluorescent protein fusion expression in organisms or cells in which...toolbox of genetic methods, such as Tol2 transposons, Cre-mediated site-specific recombination, morpholinos...

  38. Viral Production

    Type
    Collection
    ... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core. Transfections...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Transducibility Some viral vectors are tested in vitro and in vivo for gene expression and/or function. These....1E6 vg/mL, or both. Two days later, Cre-dependent GFP expression was detected with direct fluorescence...Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...

  39. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... Empty Vectors Many gRNA empty vectors have been deposited at Addgene. To find the gRNA vector for your...constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 ...size and allow for expression of more sgRNAs in a single vector. sgRNAs are expressed as polycistronic ...gRNA expression vectors for insertion of custom gRNA target sequences. CRISPR...CRISPR Plasmids: Empty gRNA Expression Vectors Browse CRISPR Plasmids By Function Genome Editing Cut Base...BbsI none S. pyogenes Gersbach pUC57-sgRNA expression vector 51132 Mammalian BsaI none S. pyogenes Huang...Multiplex gRNA Vectors Several systems have been developed to generate multiplex gRNA vectors. These systems...

  40. Retrovirus Plasmids

    Type
    Collection
    ...63704 pRetroX GFP T2A Cre CMV/MSV Tetracycline inducible expression of GFP T2A Cre fusion in mammalian ...Ecotropic MLV expressing envelope plasmid Nienhuis and Salmon 35616 pCAG-VSVG Envelope VSV-G-expressing envelope...GFP MoMLV U6-driven plasmid for shRNA expression; also expresses GFP. See pMKO.1 puro for Puro resistance... to eGFP expression Clevers 18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression; also expresses... 9044 pMIG MSCV Plasmid for transgene expression; also expresses GFP. Also see pMIG-w , a variant that...27995 TtRMPVIR CMV/MSV Tet-regulated expression of shRNA; expresses rtTA. See Zuber et al. 2011 for additional...pLncEXP CMV/MSV lncRNA expression plasmid Sun 60683 pLXIN-Luc MoMSV Stable expression of luciferase in mammalian...
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