Skip to main content
Addgene

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

In Category

We narrowed to 919 results for: IRE;

Showing: 521 - 560 of 919 results

  1. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...effectively utilize the host cell’s inherent homology directed repair (HDR) process following the double strand...break and introducing the tag through homology directed repair To introduce the fluorescent tag to the...on the Addgene Blog Learn More about Homology Directed Repair Learn More about Non-Homologous End Joining...

  2. CRISPR in the Clinic

    Type
    Blog Post
    ...Systemic therapies involve CRISPR-Cas delivery directly to the patient. Patient-derived cells can alternatively...the goal of ‘fixing’ mutations or introducing a desired edit, but this particular class of treatments has...inflammatory diseases, blindness, and diabetes. Directions and obstacles for future CRISPR therapies CRISPR...

  3. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...longer incubation times and didn’t necessarily require cycling. The Pros and Cons Researchers used the...select for correctly assembled phage genomes, it required a very stringent selection process and benefited...selection, failed under normal cycling protocols, and required a 48-hour incubation at 37 °C. While the 52-piece...

  4. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ...Targeting Receptor Expression Using chemogenetics requires successfully introducing receptors to target cells... to control ion flux. The system allows you to directly manipulate ion channels. When working with PSAM...Magnus C, Lee P, Bonaventura J, Zemla R, Gomez J, Ramirez M, Hu X, Galvan A, Basu J, Michaelides M, Sternson...

  6. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...Development Dynamics Database (WDDD) The WDDD aims to acquire quantitative data on all the cell divisions of ...Kit for CRISPR Cas9 edits At Addgene, we have the entire SapTrap toolkit from the Jorgensen lab. SapTrap...Project: The Complete Gene Expression Map of an Entire Nervous System." Neuron 99.3 (2018): 430-433. PubMed...

  7. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...active sites. In addition, Casɸ has a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is... is expected to contribute to experiments that require high photostability, such as time-lapse imaging...Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a reduced scale...

  8. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ..., comprised of PAS and GAF domains of iRFP713. Requires presence of heme oxygenase for chromophore formation...miRFP709 and miRFP670, pair with common PAS domain. Requires presence of heme oxygenase for chromophore formation...BiFC interaction is typically irreversible and requires some time for chromophore maturation, splitFAST...

  9. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...that the orientation of the flanking loxP sites directs the Cre recombinase to invert, translocate, or ... the antisense orientation (Figure 1, top). The entire DNA cassette would be flanked by two pairs of target... Wendling O, Chambon P, and Ghyselinck NB. A directional strategy for monitoring Cre-mediated recombination...

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

    Type
    Blog Post
    ...world. With three Nobel Prize winners, one NIH Director and a co-founder of BGI-China presenting, it’s... discoveries and moving beyond DNA Eric Green, Director of the National Human Genome Research Institute...proteins). The chemistry is complex and this work requires an interdisciplinary team.  Day two of the conference...

  11. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...deposited their plasmid with Addgene, hoping to inspire others to explore the phenomenon, improve the tool...Evolution of a new variant Maria and the team set up a directed evolution and screening experiment with three ...topics. He embraces open science, self-publishing directly on his website (and often posting updates on social...

  12. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ...This will make it difficult to properly gate your desired cell population. By compensating between fluorescent...other corners of the dot plot, compensation is not required.  In Figure 3, we also know that on most cytometers... cytometers, FITC and APC should not require compensation between each other. We can therefore conclude...

  13. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...level. The heartbeat – a muscle contraction – is directly related to the electrical impulse of the cardiac... to measure membrane potential and can also be directed to specific types of cells, such as neuron subtypes...For in vivo imaging applications, the ability to direct your voltage sensor selectively is especially essential...

  14. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ... minor pre-recombined population that does not require the recombinase for transgene expression. Once ... of quality control is recommended but not yet required for clinical-grade materials, and is not commonly...https://doi.org/10.1089/hum.2019.277 Lecomte E, Tournaire B, Cogné B, Dupont J-B, Lindenbaum P, Martin-Fontaine...

  15. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...of antibiotic-resistant cells; during the time required to grease, place and fill a dozen cloning rings...endocrine cell lines we most commonly work with, require extra serum to survive at low densities - adding...growing cells such as HEK and CHO do not seem to require this, while nearly all endocrine cells do. While...

  17. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...handle or direct with a degree of skill, To work upon or try to alter for a purpose and To direct the professional... these tasks before I was a manager! Second, my direct report was not engaged because I only left him/...

  18. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...syndrome coronavirus 2 (SARS-CoV-2), the virus may acquire mutations to escape these therapeutics. Jesse Bloom...the antibodies were able to immunoprecipitate the entire complex in three different cell lines. The Gräslund...enzyme. Compared with previous research, which required protein compartments to co-localize multiple enzymes...

  20. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...alternative approach to NHEJ is to utilize the Homology Directed Repair (HDR) to knock in corrective genes. For...to repair the double strand break via homology directed repair and you would probably need to use a two...human cells being subjected to scarless homology-directed genome editing (Maggio et al., 2014). Gonçalves...

  21. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...single 96-well plate for HDR that satisfy all requirements. To alleviate this bottleneck, we began building... your own purposes, or delete the originals if desired. It works as a whole or in part. For example, you... were optimized for HDR, except for TagIn. HDR requires the additional step of mutating guide sequences...

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

    Type
    Blog Post
    ...suppressors and upregulation of proto-oncogenes ­­­can directly inhibit the ability of a cell to undergo programmed...that a number of novel small molecules can bind directly, albeit weakly, to K-Ras and inhibit its activation...A single mouse click on a gene of interest will direct the investigator to a list of all available Addgene...

  24. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...guest blogger Melanie Fox, founder and executive director of Central Indiana Science Outreach and a Postdoc...as a student. They didn’t always know the best direction to guide me because they had no experience with...Melanie Fox is currently the founder and executive director of Central Indiana Science Outreach and a Postdoc...

  25. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair (HDR) template...straightforward. Likewise for step 4, as this ties directly back to your specific goal. When thinking about...

  26. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...mammalian antibiotic selection gene. However, it does require a slower transformation recovery period than amp...zeocin resistance by expressing a protein that directly binds to zeocin, preventing it from binding DNA...low rate, there is evidence that zeocin is never entirely detoxified, even when Sh ble is expressed (Trastoy...

  27. An Introduction to Adenovirus

    Type
    Blog Post
    ...sense of what AdVs are about – and we hope that it inspires further use of existing vectors, as well as research...response to adenoviral vector infected cells does not require de novo viral gene expression: Implications for...1996). The adenovirus death protein (E3-11.6K) is required at very late stages of infection for efficient...

  28. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...dCas9, require Zinc-finger or TAL proteins targeting a specific genomic locus, which can require multiple...genomic region was an intensive process. One method required insertion of a recognition sequence into the genomic...

  29. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...big science,” a massive, expensive project that required collaboration from scientists around the world... undermining science are even more immediate and dire than Zimmer could have imagined. Does this detract...about CRISPR alone, and Zimmer himself has written entire books about optogenetics. Defining science and ...

  30. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...to ddPCR. Droplet digital PCR (ddPCR) does not require a standard or a reference, meaning saving on reagents...20,000 droplets. Each droplet contains the material required for a mini amplification reaction to take place...for PCR experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and Eriko Uchida. "2D droplet...

  31. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...Molecular cloning requires some method of screening colonies for the presence of an insert. Traditionally...either briefly boiling the sample before use or by directly adding a small volume of the sample to the PCR...tell you whether the insert has the correct directionality. Verifying the insert sequence with Sanger ...

  32. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...from long written proposal narratives. Now they require grant seekers to use online applications with significant...to and proof of compliance with the grantor’s requirements as well as state and federal regulations. A ... worked for an organization in a position that required some involvement in the grants process and managing...

  33. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...exposed even one time. Doctors and researchers don’t entirely understand why some people develop the disease...one of the best ways to improve the traditionally dire prognosis for mesothelioma patients is to catch ...inhibitor) prevents the growth of new blood vessels required to sustain growing tumors. Studies have shown ...

  34. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral production...efficiency can vary across cell types and generally requires optimization of many variables, such as seeding...Using pooled libraries can be intimidating and require a large amount of experimental work, but with this...

  35. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...generating reliable results from CRISPR edit data requires the help of robust software tools. As a consequence...of the edited sample. You can also download the entire analysis as a ZIP file by clicking the Download...the inferred distribution of indel sizes in the entire edited population of genomes. Hovering over each...

  36. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ... sites. In cases like these, antibodies can be directly conjugated to agarose or magnetic beads with commercially... commercially available chemical agents. Directly conjugating the primary antibody has the additional ... particularly advantageous because they do not require centrifugation between washes which can cause loss...

  37. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...on pEcgRNA plasmid as only two 24 nt oligos are required and the BsaI linearized pEcgRNA backbone can be... in large amounts and frozen for future use or directly ligated with the annealed 24 nt oligos to generate...the above systems are great for experiments that require only single chromosomal integration (ex: genome...

  39. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    .... Cre-dependent expression of the SPH activator paired with AAV sgRNA delivery make the SPH mouse an attractive...neuroscientists, not all neuroscience applications require such optimized tools. This is elegantly demonstrated...., 2018). The pooled nature of this experiment requires fluorescence activated cell sorting (FACS) to ...

  40. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...recombinase recognizes FRT sites and initiates site directed gene recombination between those sites. FRT sites...with low concentrations of vectors expressing the required recombinase. The high concentration of FLEx vectors...PMC41457. 8. Schnütgen, Frank et al. 2003. “A Directional Strategy for Monitoring Cre-Mediated Recombination...
Showing: 521 - 560 of 919 results