Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 369 results for: Cre expression vector

Showing: 201 - 220 of 369 results

  1. Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource

    Type
    Blog Post
    ...Bradbury et al., 2018), and hybridomas can lose gene expression of the mAb-encoding genes, die, or fail to revive...research materials, such as plasmids and viral vectors. However, it typically takes a few years for a ...antibodies are not monospecific: Hybridomas frequently express additional functional variable regions. mAbs 10...these issues by creating immortal antibody-producing cell lines (hybridomas) that secrete antibodies in ...Neurotechnologies® (BRAIN) Initiative, will allow Addgene to create a much needed open-access recombinant antibody/...School of Medicine on this grant. Dr. Trimmer has created thousands of high-quality antibodies/nanobodies...reagent batch. To supplement this, Addgene will be creating a crowdsourced Data Hub (built off of the concept...

  2. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9 Toxicity in Human...genome editing, activating gene expression, repressing gene expression, and much more. But there’s one... means they can be delivered in vivo using AAV vectors. Having a way to shut off CRISPR-Cas systems prevents... off-target effects Prolonged Cas activity can increase the chance of off-target editing particularly ...

  3. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    ...genes is from Proteobacteria to mammalian expression vectors. And more broadly, their data revealed that...a distantly related organism, the gene may not express well because of these different codon usage biases...closely resembles that of the host organism, it may express protein more efficiently. To classify gene sequences...rare codons, sometimes intentionally modulating expression in ways we don’t quite understand yet,” Kunjapur...synthetic. They also found out that the most common expression system within the Addgene repository is mammalian...series of simulations and machine learning, they created a classifier that predicts whether a gene was natural...

  4. Hot Plasmids - November 2023

    Type
    Blog Post
    ...fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters...reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro applications...the Wu Lab is working to create a three-dimensional atlas of protein expression in the adult mouse brain...activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive element...enable the creation of CAR-T cells directly within the body. Overall, DIRECTED can increase on-target .... The Ting lab recently made two deposits that increase the utility of this technique.  By engineering... O’Neill The Cohen lab and colleagues recently created a system for recording the transcriptional history...

  5. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...can be expressed in a single vector, pCRISPRainbow-DONOR1, with dCas9 supplied by a separate vector. ...CRISPRainbow gRNA will permit labeling, but a newly expressed standard length gRNA would “switch” the system...scaffold to enhance complex assembly, thereby decreasing the amount of background fluorescence from unbound...CRISPR to label multiple loci in distinct colors. To create a colorful Cas9 toolbox, Ma et al. turned to SpCas9...was fused to a different fluorescent protein to create three distinct colors. The specificity of these...distinct hairpin sequences produces white light. Created with BioRender.com. With this rainbow of colors...combination, for example, with a far-red FP, would increase the available colors to 15! They also envision...

  6. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...interest. In our studies, we used AAV viral vectors to express hM4Di in neurons of rhesus monkeys or mice...successful expression of the protein of interest, an alternative could be bicistronic expression of the fluorescent...the fluorescence expressed in the neuropil conceals identification of opsin-expressing cells). For other...proteins provide a versatile method to identify the expression of transgenes of interest. Furthermore, fluorophore...can be a roadblock, rather than an aid, in the expression of the protein of interest. The green fluorescent...’ tags to identify proteins Given the limited expression of hM4Di‐mCherry at the plasma membrane in monkey... mCherry with the HA tag resulted in abundant expression of hM4Di in plasma membranes in both species ...

  7. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...instance, if you were cloning a gene into an expression vector, you would want the start of the gene to end...Your Insert and Vector by Gel Purification     Now that you’ve cut your insert and vector, unfortunately...unsuccessful 1), in a vector alone plate with many more colonies than the vector + insert plate (unsuccessful... ligation step. 3. Ligate Your Insert into Your Vector In the ligation step, you mix your purified, cut...Resources on Addgene.org Find the Perfect Empty Vector for Your Cloning Experiments Browse Our Molecular... 2 ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert...

  8. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    ... compared to the E. coli strain expressing the empty vector. And thus was born ELSA - our happy lil’ cold-tolerant...decided to focus on a special class of enzymes expressed by psychrophilic (cold-loving) bacteria. These...recombinant psychrophilic enzymes for the host The expression of recombinant proteins for industrial or research...mesophilic, for example E. coli, carrying a plasmid expressing a protein of interest). Mesophilic microbes are...enzymes face very similar issues when forced to express in mesophilic host systems. The so-called “regular... to grow at low temperatures suitable for the expression of psychrophilic proteins. The primary problem...Factor), that would allow the stable folding and expression of recombinant psychrophilic proteins. As the...

  9. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    ...years, he and his colleagues built a Molecular Expressions collection including some 3,300 plasmids along... Baird said. “They are all designed in the same vector backbone with very similar restriction enzymes....shutter its doors. “We wanted to give all of these vectors to the scientific community and contribute in a...proposition, explained Michelle Baird, who has created plasmids as a technician in the Davidson lab for...prompted him to deposit the tools he and his team created with the research community at large, "[Fluorescent...

  10. Hot Plasmids - February 2022

    Type
    Blog Post
    ...individually expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins ...through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either 10, 30, 50...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure... Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when encoding multiple guides. While...opportunity for lab members or students new to protein expression and purification. Figure 3: The Penn State...Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins...expanded the genetic screening toolkit by optimizing a Cas12a-based method for screens. Cas12a’s ability ...

  11. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...excellent start (#22988, #137884). Both vectors allow the creation of amiRNAs based upon miRNA precursors...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region...considerations, such as the starting material (DNA-based vector or purified RNA) and the initial state of the dsRNA... easiest approach to induce RNAi involves the expression of long hairpin RNA (hpRNA). This method is facilitated...construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene target sequence... Although downstream work is required, several vectors can be found at Addgene that could serve as an ...ones? Here, I’ll cover some screening methods to identify them. Visual screening Regardless of the RNAi-trigger...

  12. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...then easy to package in low-capacity vectors like AAV, a popular vector due to its low immunogenicity. But...in size and are easy to package in low-capacity vectors! These small, but mighty type VI-D enzymes are ...collateral cleavage does not occur when Cas13d is expressed in a mammalian system. Since Cas13d is functionally...RspWYL1 increases both targeted and collateral RNA degradation by RspCas13d. RspWYL1 also increased EsCas13d...Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR repeat array near a putative... WYL-domain-containing accessory protein WYL1 increases RspCas13d and EsCas13d cleavage activity The majority...

  13. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...gene expression. The regulatory element is cloned upstream of the luciferase gene, and expression of luciferase...backbones with the luciferase gene, a variety of expression vectors, and literally thousands of reporter constructs...wavelength of light they emit or to optimize their expression in mammalian systems. Our luciferase collection... AkaLumine ATP No Common uses for plasmids expressing luciferase Luciferase reporter assays for monitoring...luciferase assays have become a gold standard in gene expression analysis to investigate the effect of regulatory...interactions Split luciferases, such as NanoBiT, are expressed in two subunits and only catalyze a luminescent...reaction catalyzed by luciferase, releasing light. Created with BioRender.com. Bioluminescent reporters...

  14. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...in a single vector. In addition, the pol III promoter is relatively weak, and low expression of gRNAs from...III transcription, potentially increasing the levels of gRNA expression. Since tRNAs are among the most... confirming gRNA cleavage, they examined gRNA expression levels and indel generation for two PTG constructs...protoplasts. Compared to matching traditional gRNA-expressing constructs, transcript levels of PTG1 and PTG2...Xie et. al are specifically designed for plant expression, PTGs will likely be adaptable to many different...capacity. Still confused about the various tRNA expression strategies? No worries, I've summarized the three...research? Check out the PTG plasmids or plant expression systems available from Addgene. If you have any...

  15. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...Model Organisms Plasmids 101: Yeast Vectors Tips for Screening with the Yeast Two Hybrid System ...Phillips made the decision to release the K. phaffii expression system to the general research community in the...different uses for the phaffii-lous strain. Protein expression   K. phaffii first gained attention for its powerful...This gene promoter, AOX1, was co-opted for the expression of recombinant proteins by the pharmaceutical...pharmaceutical and biotech industries as a protein expression workhorse. This yeast can also give more protein bang...Saccharomyces cerevisiae to human: The important gene co‑expression modules. Biomedical Reports 7.2 (2017): 153-158...protein secretion, making harvesting recombinant proteins a cinch. This is because it doesn’t secrete its ...

  16. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...viral vectors.  Caspedia: A data repository for Cas enzymes.  Open repository of CRISPR screens Cellosaurus...enzyme cut sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled... sequence and functional information, such as expression patterns, post-translational modifications and...plasmid analysis tool including editing, annotating, creating maps, and more. This tool is maintained by M. ...University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid pET21a-BirA. ...

  17. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...interest: Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial Expresses Aquamarine with N-terminal...Cyan Mammalian Expresses mammalian optimized Aquamarine mCerulean N1 Cyan Mammalian Express a gene of interest...Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial Expresses CyPet with C-terminal... Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial Expresses YPet with C-terminal...Mammalian Express a gene of interest fused to the C-terminus of SYFP2 Clover Green Mammalian Expresses Clover...Cyan & Yellow Bacterial Gateway-compatible vector to express a gene of interest fused between ECFP and ...monomeric Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric...

  18. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...of robust expression of high G+C content genes, as validated through monitoring the expression of fluorescent...different vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check... be seen through the success of the change in expression of GFP, for the stop codon should have changed...different biochemical secondary energy sources to increase protein yields from 36 μg/mL to 266 μg/mL. The...oxytetracycline enzymes. Next, this system was used to recreate natural biosynthetic pathways. Biosynthetic pathways...

  19. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...Non-autonomous transposons, however, need another TE to express a transposase or reverse transcriptase so they ...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ... in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase,...efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold...transfer Transposons are an alternative to viral vector delivery of transgenes, like for iPSC reprogramming... piggyBac, which is a big advantage over viral vectors (~5 kB payload for AAV and ~8 kB payload for lentivirus...

  20. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ...plant We developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome...genome. We constructed three versions of the vector (pKABE6.3, pKABE7.9, and pKABE7.10) so that the user...are better suited (Gaudelli et al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA...scaffold) in a pol-III (OsU3) promoter driven expression system. We use a single polycistronic tRNA-gRNA... genomic site simultaneously.     These binary vectors could be used to generate stable genome edited ...Agrobacterium-mediated transformation. We also constructed smaller vectors suitable for transient assays using protoplast ...cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates a nick in the DNA by cutting only a single strand...
Showing: 201 - 220 of 369 results