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We narrowed to 430 results for: CAPS;

Showing: 81 - 120 of 430 results

  1. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...terminus of Cas9 H840A nickase. The fusion enzyme is capable of installing targeted insertions, deletions, and...

  2. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...nickase fused to a reverse transcriptase (RT) is capable of installing targeted insertions, deletions, and...

  3. Kazuhiro Oka Lentiviral Vectors

    Type
    Collection
    ... WPRE has been removed to increase the cloning capacity. pCDH-CB-IRES-copGFP-T2A-Puro 72299 Express gene...

  4. CRISPR Plasmids - Plants

    Type
    Collection
    ...nickase fused to a reverse transcriptase (RT) is capable of installing targeted insertions, deletions, and...

  5. Lentivirus Plasmids

    Type
    Collection
    ...envelope plasmids) encode components of the viral capsid and envelope and are used in conjunction with the...

  7. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...vectors have both distinctive capsids and genomic structures. The viral capsid is the key that lets AAVs ...of promoter/enhancer. Table 1: AAV capsids and receptors. Capsid Primary receptor Coreceptor AAV1... true! In a previous post, we covered systemic capsids, which allow AAVs to broadly spread throughout ...particular cell type depends largely on the viral capsid, but specificity doesn’t end there: expression ...engineered serotypes by mixing, matching, and modifying capsid and genomic structures. For example, rAAV pseudotypes...pseudotypes mix the genome of one AAV serotype with the capsid of another. Formally, pseudotypes are described...refers to the genome, and the second number to the capsid. Most vectors provided by Addgene are pseudotypes...

  8. Rett Syndrome

    Type
    Collection
    ... MECP2 null alleles or common mutations that recapitulate Rett syndrome in humans. MECP2 knockout rats...

  9. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...-enveloped particle of ~100 nm, consisting of a capsid surrounding an inner core which contains the adenoviral...~ 100nm in size, and consists of an icosahedral capsid made of 3 types of proteins: fiber, penton and ...protected by histone-like proteins forming the nucleocapsid. B) Three generations of adenoviral vectors:... Ad vectors (helper-dependent (HD-Ad), or high-capacity (HC-Ad__ are devoid of viral genes and therefore...such as HEK293 cells. 1st generation AdV cloning capacity is limited to 8.2 kb, and in vivo transgene expression...Gen AdVs were designed to have increased cloning capacity, decreased  potential for the generation of RCA...2nd generation AdVs, thereby increasing cloning capacity to ~12 Kb. The ever popular pAdEasy system belongs...

  10. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...in this post: Pooled library to study antibody escape in SARS-CoV-2 Recombinant antibodies for aminoacyl...New ready-to-use viral preps Mapping antibody escape in SARS-CoV-2 by mutational scanning By Seth Kasowitz...SARS-CoV-2), the virus may acquire mutations to escape these therapeutics. Jesse Bloom’s lab at the Fred...From Starr et al Cell., 2020. (B) Mapping antibody-escape mutations with the yeast display library. From ...rate-limiting enzymes. In this method, the major capsid protein VP1 is co-expressed with a fusion protein...VP1, which self-assembles into a compartment encapsulating the protein of interest (Figure 3).   ...D-glucaric acid biosynthesis. This MPyV system encapsulated a rate-limiting enzyme in this pathway (myo-...

  11. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...process of anchoring an antibody, often called the capture antibody, to agarose or magnetic beads in a way...immunoglobulin, leaving the antigen binding sites free to capture the target protein. The beads can be collected ..., and perform the IP at 4oC. When choosing the capture antibody for IP, be sure to choose one that recognizes...proteins that may interact nonspecifically with the capture antibody or Protein A/G. To preclear, incubate ...nonspecific antibody from the same host species as the capture antibody. This will deplete the nonspecific proteins... the target protein in the sample binds to the capture antibody and is immobilized. The ideal antibody...antibody concentration to use will vary between capture antibodies and can be determined through titration. ...

  12. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...relative to the total number of viral capsids (which can include empty capsids that are devoid of a viral genome... viral particles. Ratio of full to empty viral capsids: the ratio of genome-containing viral particles...genome). Empty viral capsids can be formed during viral production depending on the  experimental conditions...One study showed that approximately 50% of viral capsids were genome-containing (“full”) (Zeltner et al....1999). The percentage of genome-containing viral capsids is typically quantified by electron microscopy ...we are pleased to report that over 95% of viral capsids in representative viral vector preparations are...purified viral vector solution (which contains viral capsid proteins) as opposed to purified DNA. This may ...

  13. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...and invade multiple organs, and most importantly escape the immune mechanisms of the host. The cause, manifestation...drug resistance in bacteria, the tumor cells that escape the elimination stage acquire genetic alterations...antigen production to evade the immune system. 3. Escape - Tumor cells that survive this phase are able ...outsmart the host immune mechanisms altogether and escape future immune surveillance. Dormancy of certain...interactions with the immune system, tumors “learn” how to escape it. In this process, cancer cells develop and thrive...Evading Recognition Tumor cells over-express CD47 to escape recognition by circulating macrophages. Tumor cells...metastasis, block the cell-surface proteins that help escape immuno-surveillance, and curb cytokine and chemokine...

  14. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    ... users coat a multiwell plate with a capture reagent. The capture reagent may be an antigen or an antibody...on how the assay is set up. Then they block the capture reagent to prevent non-specific interactions. Next...ELISA the multiwell plate is first coated with a capture antibody that binds to the antigen in a sample ...ELISA, users first coat a multiwell plate with a capture antibody and then add their antigen-containing ...antigen-containing sample. The target antigen binds to the capture antibody and a series of wash steps removes non-target....  Importantly, the detection antibody and the capture antibody must bind to distinct, non-overlapping...they require two independent antibodies for the capture and detection steps, which increases the specificity...

  15. Zhang Lab CRISPR Page

    Type
    Collection
    ...(Cas) genes as well as non-coding RNA elements capable of programming the specificity of the CRISPR-mediated...

  16. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...developing new capsids through direct evolution, capsid shuffling, and peptide displays. These capsids will be...slight variations in their viral capsids. Novel forms of AAV capsids are discovered frequently and recent...activation of innate immunity. AAV vectors are also capable of infecting a wide range of cell types including...enabled us to generate new and refined forms of capsids with improved specificity. AAV vectors are able...

  17. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...be packaged in a variety of capsids. To date, at least 12 different capsids, or serotypes, have been identified...using AAV is the limited packaging capacity. Lentiviruses have capacities of 8–12 kb, while AAV only has ...libraries available at Addgene. The large packaging capacity and stable expression of lentiviral vectors make... AAV, with the genome from one serotype and the capsid of another. Researchers have used these psuedotypes...Interest in interrogating and mapping the genetic landscape of tissue-specific diseases has grown exponentially...

  18. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...antibodies, Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17] and Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86], target...target distinct epitopes on the SARS-CoV-2 nucelocapsid protein, one of the virus’s most abundant proteins...Figure 3: HeLa cells transduced with SARS-CoV-2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane...) were immunoblotted against Anti-SARS-CoV-2 Nucleocapsid Protein antibodies #211756 (left panel) or #211757..., et al. (2021). Development of a SARS-CoV-2 nucleocapsid specific monoclonal antibody. Virology. 558,...-strand AAV (ssAAV). To maintain the packaging capacity and improve the specificity of ssAAV, a targeting...

  19. Bacterial Expression Systems

    Type
    Collection
    ...tuberculosis Vinay Nandicoori 78577 pL99 PnitA-NitR ε-caprolactam Escherichia coli , Streptomyces sp. Xuming Mao...

  20. CRISPR Pooled gRNA Libraries

    Type
    Collection
    ...56,762 (Set A) 56,476 (Set B) Broad GPP activation Caprano p65-HSF 92383 (Set A) 92384 (Set B) Activation ...

  21. Early Career Researcher Toolbox: Free Tools for Making Scientific Graphics

    Type
    Blog Post
    ...intuitive and I was making figures quickly. Inkscape Inkscape is the most similar to Adobe Illustrator out...and like Vectr, Inkscape uses vector graphics. There’s a steep learning curve for Inkscape, but there are... desktop use for Windows, Mac OS X, and Linux. Inkscape is versatile and has a wide selection of tools...lots of resources for learning the software. The Inkscape website has many tutorials and Lynda has a tutorial...

  22. Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants

    Type
    Blog Post
    ... compared to the wild-type, escaped neutralization. “That degree of escape from a very small number of...luminescence.     B.1.351 variant neutralization escape The lab saw that the B.1.351 variant was very resistant...what was surprising was the extent to which the escape appeared to be happening.”  In their assay, the...-YS, Lessells RJ, de Oliveira T, Sigal A (2021) Escape of SARS-CoV-2 501Y.V2 from neutralization by convalescent..., Balazs AB (2021) Multiple SARS-CoV-2 variants escape neutralization by vaccine-induced humoral immunity...

  23. Hot Plasmids - May 2022

    Type
    Blog Post
    ... AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids are ...positive and negative selective pressures for novel capsids. In a continuation of that work, they have found...is true in both mice and nonhuman primates (for AAV.CAP-B10, for example).  Given that the liver is an ... derived from AAV9.  Of those shown here, only AAV.CAP-B10 causes brain wide expression, with no liver...

  25. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...to the type of capsid (outer protein shell) used to package an rAAV vector; these capsids allow the virus...AAV components      Cargo capacity Empty rAAV has a cargo capacity of ~ 4.7 kb, meaning that your... virus, packaged in a non-enveloped icosahedral capsid, that can be used to express genes of interest ...plasmid and individually packaged into the AAV capsids. As a genome conversion in the transduced cell ...

  26. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...select novel AAV capsid variants could unintentionally limit the utility of candidate capsids.  Are you interested...small randomized peptide loop inserted in the AAV9 capsid. This mutant library was then screened in vivo ...version of LY6A, but a mutation in PHP.B’s unique capsid loop disrupted binding. Collectively these results...LY6A, a novel cellular receptor for AAV-PHP. B capsids." bioRxiv (2019): 538421. If you’re interested ...administration of the adeno-associated virus-PHP. B capsid fails to upregulate transduction efficiency in ...

  27. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ... determine if CRISPR/Cas9 escape occurs in an animal model. If these escape mutations are frequent, the...their results, Wang et al. showed that HIV-1 could escape from CRISPR/Cas9-induced modification targeting...either the LTRs or essential genes. Many of these escape mutations were located near the Cas9 cut site, ...1 infection, which would make it easier for an “escape” virus to propagate and infect adjacent cells. ... Inhibit HIV-1 Replication and Accelerate Viral Escape. Cell Rep. 2016 Apr 5. PubMed PMID: 27068471. Ramanan...

  28. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ... for the identification of plasmid multimers. Capillary Gel Electrophoresis (CGE) analysis Another technique...technique that can detect multimers is capillary gel electrophoresis (CGE), which has been used in plasmid...Thompson L. Vaccine Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med. 2020;20(10)...PMC4480603. Holovics HJ, He Y, Lacher NA, Ruesch MN. Capillary gel electrophoresis with laser-induced fluorescence...fluorescence of plasmid DNA in untreated capillary. Electrophoresis. 2010 Jul;31(14):2436-41. doi: 10.1002/...study and topology analysis of plasmid DNA by capillary gel electrophoresis. BioProcess Int. 2006 Sep;...

  29. Antibodies 101: Validation

    Type
    Blog Post
    ...your experiments. Capture MS Strategies This approach uses immunoprecipitation to capture proteins from a...and then uses mass spectrometry to identify the captured proteins. If the antibody is specific, then you...approach is ideal for validating antibodies for immunocapture applications and is amenable to high-throughput...to validate that your antibody is specifically capturing your RNA-binding protein of interest. So you ...immunoprecipitation with your antibody and send the captured proteins off for mass spec analysis (above). When...

  30. Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight

    Type
    Blog Post
    ...distinct behaviors such as spontaneous swimming and escape reflex. These traits, coupled with short generation...zebrafish at 5 days post fertilization, including escape behavior and spontaneous swimming. When startled...zebrafish react by performing a distinct C-bend to escape possible danger (see figure). Larvae also perform...in response to any specific stimulus. While the escape response neural pathway is well characterized, ... kind of swimming behavior, spontaneous swim or escape, was elicited when these different neuron groups...

  31. An Addgene Summer Internship

    Type
    Blog Post
    ...my training, I dove straight into my capstone project. A capstone project is a major project given to ...Figure 1: Addgene's summer interns presenting their capstone projects during the Summer Intern Poster Presentation... scientist. But I expected something wrong. My capstone project was based on plasmid cloning and I, the... Once everything was confirmed, I worked on my capstone project report and presentation, during the final...

  32. Antibodies 101: ChIP

    Type
    Blog Post
    ...interactions. ChIP works by using antibodies to capture protein:DNA complexes with antibodies specific ...your protein(s) of interest. Once the complex is captured, it is treated so that the protein and DNA are...DNA and then determine concentration.  Protein Capture After crosslinking and sonication or enzymatic ...antibodies, with your samples, and then use an antibody capture method such as Protein A/G beads to collect your...and the Three Steps (Crosslinking, Fragmenting, Capturing), if you ask me.  Each of the three steps need...

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

    Type
    Blog Post
    ...the potential hazards of creating “gene drives” capable of spreading through wild populations. Whereas ...organism carrying a synthetic gene drive were to escape the laboratory, the drive could eventually spread...protocols, they can reduce the risk of accidental escape to a negligible level. One major reason we chose...chose to publish the concept and likely capabilities of RNA-guided gene drives in the summer of 2014 (after...performing experiments in a geographic area where escaped organisms won't be able to find mates2. For example...

  34. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...polymerase (reviewed in 2, 5). The minigenome is capable of being replicated and transcribed in the cell... Minigenome System). However, minigenomes are incapable of producing infectious virus (similar to lentiviral...VP30, and L (reviewed in 5). The RNA genome is encapsidated by the nucleoprotein, NP. NP associates with...mRNAs are translated) by cellular machinery. NP encapsidates the negative sense, single-stranded RNA minigenome...viral polymerase complex then recognizes the encapsidated minigenome as a template for replication and...

  35. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids. Human Gene Therapy Methods 27...make it a great viral vector, but its packaging capacity is limited to ~4.7kb, or roughly half the packaging...Lastly, depending on its design, donor A may be capable of expressing a truncated protein. This can be ...Expands Adeno-associated Viral Vector Packaging Capacity in a Transgene-independent Manner. Molecular Therapy...

  36. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...methods termed proximity-based labeling to better capture these interactions in a vast array of biological... of biotin for avidin, proximity labelling can capture weak/transient protein interactions as well as ...exposed lysines on neighboring proteins. Thus to capture protein complexes, scientists can simply express...After a few hours biotinylated proteins can be captured on streptavidin affinity beads or matrices. The...transpeptidase sortase A variant (mgSrtA), which is capable of promiscuously labeling various cell surface ...

  37. New FLIS Plasmid (April Fools 2022)

    Type
    Blog Post
    ...of working with fruit flies is their ability to escape their habits and live in the lab itself. Many of...of tiny black dots during our stay there. Yet capturing the flies is nearly impossible, due to their small...everywhere find and re-contain any specimens that escape during routine lab activities.  Here, we demonstrate...plasmid-based system for locating and containing escaped flies. Nature 4:1-22. https://doi.org/4.01.2022...

  38. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ... and migrate within neurons, and that are also capable of spreading across synaptic connections. Most ... like lentivirus, but instead is contained by a capsid which is comprised of the viral proteins VP1, VP2...genetically engineered to have differences in their capsid proteins. For a brief overview see Addgene’s AAV...relatively small (20 nm) in size which makes them capable of diffusing in the extracellular space; they also...include AAV1 – 9, with most of these serotypes capable of entering neurons (see table 1)(Choi et al., ...2017 also provide evidence that AAV1 and AAV9 are capable of synaptic spread. The underlying mechanisms of...

  39. What's New in CRISPR - December 2019

    Type
    Blog Post
    ...editors CRISPR toolkit for epitope tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery Cytosine...protein-truncation strategy with reduced RNA-recognition capabilities.  Read the paper in Nature Biotechnology Find...Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle Transient expression of CRISPR...Recently, Baisong Lu’s lab developed a lentiviral capsid-based bionanoparticle system for packaging Cas9...
Showing: 81 - 120 of 430 results