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We narrowed to 294 results for: expression vector

Showing: 201 - 220 of 294 results

  1. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    Published
    Jan. 18, 2018, 2:34 p.m.
    ...inducible system is found in many bacterial expression vectors. Negative inducible promoter pBad is another...gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant...Temperature inducible promoters Temperature sensitive expression systems are typically less leaky than chemically...chemically induced promoters; they show near-zero expression at regular temperatures but can be induced by ... promoters, in which a gene of choice is only expressed following exposure to a brief heat shock. In the...promoters Light is another way to activate gene expression, and two-component systems used in synthetic ...transcription from phage promoter pR, preventing expression of a reporter gene. When light is present, YFI...

  2. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...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...

  3. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ... mutations and gene expression changes elsewhere in the genome. Second, expression is by nature transient...control could enter cultured cells and activate expression of a luciferase reporter. CPP was not required...stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. CRISPR...causes a cell to take up, and continued protein expression would require multiple transduction events. For... to binary systems in which transient protein expression has a long-lasting effect, such as Cre- or Cas9...limitations of packaging size associated with viral vectors. For these reasons, iTOP, especially in tandem ...

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

    Type
    Blog Post
    Published
    May 11, 2021, 1:15 p.m.
    ...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 ...

  5. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters...to reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro applications...activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive element...to create a three-dimensional atlas of protein expression in the adult mouse brain using thick-section ...

  6. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    Published
    Feb. 28, 2017, 3:30 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    July 29, 2014, 6:16 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Jan. 28, 2020, 2:35 p.m.
    ... 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. Celebrate Open Data Day with Addgene’s AAV Data Hub!

    Type
    Blog Post
    Published
    March 5, 2020, 2:15 p.m.
    ...methods did you use for expression of AAV cargo? Did you use GCaMP expression? Direct Fluorescence? Immunohistochemistry... visualized expression after AAV injection, what region and cell class ended up expressing your virus?...Addgene.org Read our AAV guide Browse all  AAV vectors Find ready-to-use AAV preps ...

  10. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ... that expresses Cre with a strain that expresses a floxed gene of interest. Cre may be expressed globally...the appropriate primers to detect your targeting vector or gene of interest. 2. Breed your parental mice...infertility and can arise due to copy number, expression level, and site of integration of the transgene...in cell lines to determine how it impacts the expression level of the transgene. In existing strains of...heterozygous background to reduce copy number or expression level effects.  Another challenge that may arise...heterozygous for each mutated allele.  Cre-lox expressing strains Cre-lox strains are a common example ...we explained how genetic recombination in mice expressing both Cre and LoxP occurs. But how are these mice...

  11. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    Published
    Aug. 5, 2021, 12:15 p.m.
    ...working on developing new surface display vectors to improve expression and trafficking of these biosensors....gray (PAG), with red neurons expressing tdTomato and green neurons expressing cFos. Image from Elizabeth... Figure 2: Micrographs of the biosensor expressed in the human cell line, Hep3B. From left to right...opioid withdrawal with tdTomato and assessed cFos expression. She will explore the connectivity patterns of...chemogenetics to study the activity of parvalbumin (PV)-expressing neurons, neurons that the lab previously found...the receptors to monitor their localization and expression. Qiu saw that inhibition of the PV neurons resulted...response due to blue light stimulation of dual virus-expressing CR neurons. (C) Duration of painful responses...

  12. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...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...

  14. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must go through ...choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal injections...for the delivery of various therapies and viral vectors. This injection sounds very disruptive, but surprisingly... This is most often used for delivery of viral vectors, though drug treatments are also possible. One ...Addgene.org Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati...

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

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...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 ...an excellent start (#22988, #137884). Both vectors allow the creation of amiRNAs based upon miRNA precursors...

  16. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ... deliver a pooled population of gRNAs to Cas9 expressing cells. Pooled lentiviral CRISPR libraries (referred...population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within the genome...gRNAs are then cloned into lentiviral transfer vectors to create the CRISPR library. CRISPR libraries ...) to be used to generate lentivirus (B). Cas9-expressing cells or wild-type cells are treated with lentivirus...

  17. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    Published
    July 22, 2025, 1:15 p.m.
    ...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...

  18. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...in a single vector. In addition, the pol III promoter is relatively weak, and low expression of gRNAs from...transcription, potentially increasing the levels of gRNA expression. Since tRNAs are among the most abundant molecules... 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...

  19. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...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...

  20. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    Published
    April 4, 2023, 1 p.m.
    ...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...Five Popular Model Organisms Plasmids 101: Yeast Vectors Tips for Screening with the Yeast Two Hybrid ...
Showing: 201 - 220 of 294 results