Skip to main content

We narrowed to 842 results for: Eras

Showing: 261 - 270 of 842 results
  1. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    Published
    Dec. 20, 2018, 1:15 p.m.
    ...Cas9 in an attempt to create gene-edited babies. Several hours later, the scientist, Jiankiu He, confirmed...such edited babies were born in a twin live birth several weeks previously. While this claim has yet to be...public consensus on genome editing, there was a generally agreed-upon international moratorium on utilizing...with healthy mitochondria, with the intent to eradicate mitochondrial diseases. The key success of the...issues raised around mitochondrial replacement therapies. In other words, rather than having a meeting...bedside is, in addition to thorough ethical considerations, in making sure that the research has been ...simulations, in vitro research, animal models, and several stages of human clinical trials. This model also...
  2. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...corresponding sequencing quality. Generally, you want to have 100x-1000x coverage of your library: in other words...transformation and amplification protocol, but the overall workflow is similar for most libraries. Here are...of the final recovered DNA. This bias can be counteracted by growing the cells on solid plates, where ... extraction. Prepare library product for next-generation sequencing Once the DNA has been extracted, the...of the library should be confirmed using next-generation sequencing (NGS). For gRNA libraries, you’ll ...using both pre- and post-amplification DNA and generates PCR sequencing products from both. These products...post has helped clarify some of the important considerations when using, amplifying, and verifying pooled...
  3. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 8, 2019, 1:54 p.m.
    ...requested from Addgene psPAX2  - psPAX2 is a 2nd generation lentiviral packaging plasmid from the Didier ...and tet genes. It can be used with 2nd or 3rd generation lentiviral vectors and an envelope expressing...BB)-2A-GFP (PX458). lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang...founding. Find pCMV-VSV-G. pRSV-Rev - This 3rd generation lentiviral packaging plasmid from the Trono lab...lab. Find pRSV-Rev. pMDLg/pRRE - This is a 3rd generation lentiviral packaging plasmid from the Trono lab...This plasmid from David Sabatini’s lab is a 3rd generation lentiviral vector that contains a scrambled shRNA... This vector from the David Root lab is a 3rd generation lentiviral transfer plasmid for cloning and expressing...
  4. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    Published
    Nov. 5, 2019, 1:28 p.m.
    ...enhanced site accessibility In order to generate a genomic alteration at a specific location, it must be accessible...appropriate for your specific research question. Considerations when choosing your Cas nuclease When choosing... off-target cleavage should be an important consideration during any experiment. Sensitivity Cas9 sensitivity...Comic by Maya Kostman. Another important consideration when choosing a Cas9 is balancing specificity...Jennifer Doudna Labs used targeted mutagenesis to generate hyper accurate Cas9 (HypaCas9). This nuclease ...the Jungjoon Lee Lab used directed evolution to generate Sniper-Cas9 resulting in a Cas9 with improved ...accessibility, specificity, and sensitivity are key considerations in choosing a Cas protein for your experiment...
  5. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ...contributed by Jennifer Tsang AAVs are a great gene therapy delivery mechanism, but their small packaging capacity...splitting removes any exogenous sequences and regenerates a native peptide bond at the split site. ...tested the base editors in vivo and achieved therapeutically relevant editing efficiencies in the mouse ...comparable to those currently used in human gene therapy trials. They further tested the base editors by...Niemann-Pick disease type C, directly demonstrating the therapeutic potential for the split base editors. Find the...conversion of CO2 into water and electrical energy generation. Alistair McCormick’s lab recently developed ... Using parts from this kit, the McCormick lab generated knock-out and knock-in mutants, determined protein...
  6. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...assumption into question. Specifically, CRISPR-generated mutants believed to be knockouts can maintain ...Morgens et al., 2016). Only upon combining the data generated from both methods did a more comprehensive gene...Consequently, I will focus on different approaches and considerations present at the first stage of an RNAi-based... of the RNAi trigger requires a few initial considerations, such as the starting material (DNA-based vector...prevalent method due to ease of use and capacity to generate a long-term RNAi phenotype. Long hairpin RNA The...Importantly, instead of the assortment of siRNAs generated from a long hpRNA trigger, only a single ~21-nt...amiRNA). Although downstream work is required, several vectors can be found at Addgene that could serve...
  7. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...validated? This post will go over some of the general considerations you should review before using an antibody...important to know what you are aiming to achieve. In general, these are three core goals of antibody validation...different applications. Here, I want to highlight the general suggestions given by the International Working ...these pillars is that they are intentionally generalizable and not specific to certain applications or ...analysis of transcriptomic or proteomic data from several different tissues or cell types. If your antibody...the brain, but almost none in the liver (A). In general, RNA levels tend to correspond to protein levels...s important to understand if protein:protein interactions could be a factor in your analysis. Example:...
  8. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    Published
    Oct. 9, 2018, 12:21 p.m.
    ...enter postdoctoral training. Similarly, there are several factors to consider when selecting a postdoc mentor...system more, you will have a better idea of how to leverage it to ask new and interesting questions in your...confidence in the reagents and outcomes. This is preferable to finding out you've been doing something all...exploration, discovering new information. There are literally infinite questions one could pursue and experiments...the data and any recent relevant studies in the literature. Plus, presentations benefit from practice. If...better slides that more effectively tell a story? Generally, the answer to the second question is yes. As ...and how your research fits in with the current literature. Finally, if you give your talk or poster to ...
  9. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    Published
    Nov. 5, 2024, 8:32 p.m.
    ...establishment of two cell lines from the insectspodoptera frugiperda (lepidoptera; noctuidae). In Vitro, 13(4), ...packaging plasmids (three packaging plasmids for 3rd generation lentivirus production). The packaging plasmids...Insect cells are a less common option for the generation of viruses, though they are a popular choice ...from the pupa ovarian tissue of fall armyworms (Spodoptera frugiperda). Much like HEK-293T cells, they are...much larger packaging capacity — up to 38 kb! Generally, insect cell production methods produce lower ...versus insect cells for viral production Consideration Mammalian cells Insect cells Viruses AAV and...adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https://doi.org/10.1089/10430340260355347...
  10. Plasmids 101: Degron Tags

    Type
    Blog Post
    Published
    March 14, 2024, 1:15 p.m.
    ... molecule). Degron tags can range in size from several kDa up to ~35 kDa and are typically knocked-in ...manipulate protein expression. These tags offer several advantages over the alternative systems – CRISPR...interest to the proteasome for degradation. Like several other tags described here, the dTAG uses a small... and widespread usage, a proteolysis-targeting chimera (PROTAC) was developed for HaloTag. PROTACs are...their target protein and an E3 ligase through interacting ligands. After an E3 ligase facilitates ubiquitination... leakiness associated with the system, but the general concept of the system has remained the same. An...predict precisely how a given degron tag will cooperate with your protein of interest and how effective...
Showing: 261 - 270 of 842 results