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We narrowed to 750 results for: tro

Showing: 281 - 300 of 750 results

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

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ... disease. To do so, their T-cells were grown in vitro, their mutations corrected, and the T-cells were... body and thereby correct the mutations using retroviral vectors. However, many diseases require techniques... B, LPL deficiency, Cystic fibrosis, Muscular Dystrophy, Parkinson’s disease, and HIV. A notable landmark...infect target tissues. Improved specificity through control of gene expression and the selection of specific...CRISPR-AAV vectors Learn about Addgene AAV Quality Control Process Resources on Addgene.org Find ready-to-use...

  2. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...better than a nitrocellulose membrane. If you have already run your blot using nitrocellulose, you may decide...isn’t too different, consider antibody affinity: stronger binding antibodies will require harsher stripping...instructions for the membrane you are using. Dry nitrocellulose membranes are brittle, so proceed with caution...depend on your blot — antibodies that bind more strongly or that have saturated the blot may need longer...

  3. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ...Tsang Fluorescent proteins (FPs) traditionally introduced in Caenorhabditis elegans on extrachromosomal...gene into the target gene’s native locus, but introducing large genetic changes in C. elegans with CRISPR... donors (ssODN). So how can you use CRISPR to introduce large FP genes into C. elegans? By using a split...protein. They used CRISPR with a ssODN template to introduce the small wrmScarlet11 which is less than 60 nt...exclusively in huge bacteriophage genomes. It’s active in vitro and in human and plant cells. Its small size offers...

  4. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...method can be the absence of suitable controls. The best control for this type of assay is a clonal cell...using a reverse transcriptase mutant in a separate control reaction. This will allow you to quantify the amount...of the qPCR target gene. Developing the proper control line typically requires transducing with a limiting.... 3. Zhang, Bing, et al. "The significance of controlled conditions in lentiviral vector titration and...

  5. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    Published
    July 23, 2015, 2:30 p.m.
    ... our members took an Arduino microcontroller and a motor controller and made it spin again!  We’ve also...scientists can get involved in modelling, the electronics folks can build a robot worm on top of that model...instance, the centrifuge in the picture had a broken controller and would not spin. It was going to cost far ...far more than it was worth to repair the controller and a new replacement was not possible, but one of our...

  6. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    Published
    Oct. 30, 2018, 1:04 p.m.
    ...preimplantation embryos that are produced during in vitro fertilization treatments and donated afterwards ... produced Ethical concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation... the reprogramming factors were delivered via retroviral infection [8]. The work was taken forward the...generation of human iPSCs from dermal fibroblasts via retroviral and lentiviral infections respectively [9, 10...cell types. Like ESCs, iPSCs can differentiate in vitro into derivatives of all three germ layers, form ...

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

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...same result – uncontrolled movement of the vehicle or, in the case of cancer, uncontrolled cell proliferation... cellular level there is a universal theme - uncontrolled cellular proliferation. We have appreciated ...appreciated for some time the causes of this uncontrolled proliferation, in particular the roles of proto-oncogenes...opposite task, keeping cells from dividing out of control. One may compare the roles of proto-oncogenes and...

  8. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...of all distributed plasmids as part of quality control, but does not perform functional tests. The newest...produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA requires incubation at 55...can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA...does however rely on cloning into a vector or in vitro synthesis, which could be less desirable if one ...et al. "DNA-guided genome editing using the Natronobacterium gregoryi Argonaute." Nature biotechnology ...

  9. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ...qPCR assays, samples, standard curves, and normal controls must be run in triplicate.  The nice thing about... of DNA in your sample relative to your normal control - you can set up your calculations so that your...so forth. This also serves as a light technical control - an RQ value of 1.5 means you have either made... does not require a standard curve or a normal control, it can take a few hours longer than qPCR to run...AT-rich DNA and by variation in amplicon length. Am J Trop Med Hyg. 2008 Dec;79(6):887-92. PMID: 19052298. ...

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

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ... mutated gene. In a Scientific Reports paper introducing the delivery method in 2014, Manuel Goncalves...include their episomal nature and very efficient introduction of DNA into therapeutically relevant, non-transformed...B-thalassemia, Sickle Cells Disease, Duchenne Muscular Dystrophy, Alpha1-antitrypsin deficiency (most widely used... for cancer therapy. Elsevier, pp 39–67 Ding Q, Strong A, Patel KM, Ng S-L, Gosis BS, Regan SN, Cowan ...adenoviral delivery of CRISPR/Cas9. Journal of Controlled Release 298:128–141 . https://doi.org/10.1016...

  11. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...drug-resistant population compared to a non-treated control group. In this scenario, gRNAs that are “enriched...future therapeutic targets for diseases causing uncontrolled cell growth, such as cancer. Figure 2...As with any experiment, the use of appropriate controls, multiple replicates and several cell types can...of multiple gRNAs targeting the same gene can be strong evidence that a particular gene is actually important...Titering Your Lentivirus Plasmids 101: NGS Quality Control for Pooled Libraries Find More CRISPR Resources...

  12. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ... create a single polycistronic 15 kb insert. The final construct was then introduced to HeLa cells (Figure...resulting HeLa cells show robust labeling, with strong expression and detection of each fluorophore at...meaning that (in the previous example) one can control whether the third module is added to the 5’ terminus...degree of identity, so not suited to generate polycistronic containing multiple identical polyadenylation...

  13. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...techniques involve the assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation...specifically cuts methylated DNA, so it will selectively destroy the template, but not newly synthesized sequences...protein-coding sequence, or a mutation can be introduced in a gene, while adding a GFP-tag (Figure 3)....unamplifiable’ Check your amplification by gel electrophoresis. In our experience, good amplification after...interested in neuronal signal transduction, and uses electron cryo-microscopy to understand receptor function...

  14. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...diseases such as hemophilia, muscular dystrophy, spinal muscular atrophy, neurodegenerative diseases and metabolic...life-threatening (cystic fibrosis, hemophilia, muscular dystrophy)? AAV gene therapy started in the mid-90's with...larger transgenes. Unfortunately it has limited tropism in the retina, primarily infecting cells of the...epithelium (RPE). AAVs, on the other hand, have broad tropism in the eye. AAV-2, the first identified serotype...

  15. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...now many variations, including many that use auxotrophic rescue genes as reporters and some that repress...example of the former. In this system, histidine-auxotrophic host strains are used to select for binding-dependent...accounted for using appropriate replicates and controls (see Tips below), and by validating leads with...reporter gene activation. Including a prey-only control also provides a baseline level of reporter activation...that are known to bind serves as a good positive control to ensure that your setup works. The last thing...

  16. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system ...requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene...Addgene: Your lab recently developed a retroviral enChIP system using CRISPR technology. Can you describe...Associated with an IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita...

  17. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...research. Many other mutations have since been introduced to GFP and new iterations of fluorophores are... Transcription Reporter: Placing GFP under the control of a promoter of interest can be used to effectively...Scientists can use light to detect, measure, and control molecular signals, cells, and groups of cells in...Plasmids of this type may have the GFP under the control of an additional promoter from that of the gene...fate studies. It can also be used, when put under control of promoters of interest, to visualize the developmental...

  18. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    Published
    Feb. 6, 2024, 3:13 p.m.
    ...axis determined by the researcher, based on their controls. You can usually tell where this point is by looking..., M., & Al-Qahtani, M. (2018). Evaluation of in vitro chondrocytic differentiation: A stem cell research...Minguela Puras, A., Pons, J. A., & Muro, M. (2018). In vitro intracellular IFNγ, IL-17 and IL-10 producing T ..., Li, J., Greiner, D., & Anderson, D. (2015). Neutrophil Responses to Sterile Implant Materials. PloS ...Plasmid-based Recombinant Monoclonal Antibodies Introduction to Antibodies  ...

  19. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...factors. The episomal vectors are introduced into cells by electroporation. hiPSC colonies are obtained by...to perform cell therapy and to model diseases in vitro. Find stem cell plasmids at Addgene Many sources...Fibroblasts need to be cultured and expanded in vitro for several passages to produce an adequate number... by 10 days after electroporation, most of which are integration-free, karyotypically normal and are able...

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

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ...DNA dot blot...Transduction assay...SDS-PAGE...Electron microscopy… Any guesses? That's right... ways...genome-containing viral capsids is typically quantified by electron microscopy of a viral vector solution. Since this... this technique is intensive and requires an electron microscope, it is not routinely performed on all...genome-containing (Figure 1).    Figure 1: Electron micrograph of an Addgene AAV viral vector preparation...Considerations When Using AAVs Addgene's AAV Quality Control Methods Check Out Our Viral Vector Featured Topic...
Showing: 281 - 300 of 750 results