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  1. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...components, see the AAV Plasmid Elements table . Figure 2: Plasmids required for rAAV vector production. Created... pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism...the central nervous system . Molecular Therapy, 10 (2), 302–317. https://doi.org/10.1016/j.ymthe.2004.05.024...the production of AAV vectors . Microorganisms, 12 (2), 384. https://doi.org/10.3390/microorganisms12020384...retrograde access to projection neurons . Neuron, 92 (2), 372–382. https://doi.org/10.1016/j.neuron.2016.09.021... contains many viral vector-specific terms and acronyms, so if you're new to viral vectors or simply need...wild-type serotype, and a very high infectivity across a broad range of cell types in vivo. AAV-DJ has...
  2. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ..., and the desired gene (transfer vector plasmid), 2) the HIV-1 genome containing all viral genes with ...protocols that have been described (protocol 1, protocol 2, protocol 3) to find the one that will give you the...PubMed PMID: 3785217. PubMed Central PMCID: PMC367857. 2. Naldini, Luigi, et al. "In vivo gene delivery and...quiescent or terminally differentiated cells such as macrophages and microglia. This property makes the lentivirus...
  3. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    ...affinity and specificity. Trends in Immunology, 29(2), 91–97. https://doi.org/10.1016/j.it.2007.11.004 ...phagocytosis (ADCP), Fcγ receptors on macrophages bind IgG antibodies. The macrophages then engulf and digest the...antibody-dependent cellular phagocytosis for effective macrophage-based cancer immunotherapy.Sci. Adv.8,eabl9171...
  4. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...dissociation by bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha and...the potency and efficacy is typically non-uniform across the transducerome for a specific receptor-ligand...further used TRUPATH to analyze a panel of compounds across the transducerome on a single GPCR, showing the...available as a collection, facilitating screening across the transducerome, to gain a more complete understanding...
  5. Components of CRISPR/Cas9

    Type
    Blog Post
    ...Without binding to the guide RNA, Cas9 cannot cut. 2. Bind to target DNA in the presence of a guide RNA...hybrid of the tracrRNA and the crRNA. The “direct repeat region” combined with the tracrRNA forms the scaffold...which defines the genomic target for Cas9, and the tracrRNA which acts as a scaffold linking the crRNA to ...for processing pre-crRNA into mature crRNA and tracrRNA binding. The spacer regions, on the other hand..., the “direct repeat region” combined with the tracrRNA forms the scaffold portion of a gRNA and the “...and the direct repeat region. B) RNase III, the tracrRNA and Cas9, bind to these transcripts and C) cleave...cleave them leaving mature crRNAs bound to the Cas9/tracrRNA complex. D) The mature crRNA is used to guide ...
  6. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically... will likely want to monitor the voltage change across the membrane of cardiac cells. Similarly, neuronal...
  7. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...engineering methods have their own pros and cons (2). The NgAgo DNA guide is inexpensive and can be 5’...Nature biotechnology (2016). PubMed PMID: 27136078. 2. Burgess, Shawn., et al. "Questions about NgAgo." ...other hand, the widespread success of CRISPR-Cas9 across multiple model organisms (3) lies in the versatility...
  8. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...closer to the clinic (Tsai and Joung 2016). Table 2: Unbiased Genotyping Options Technique Detection...PMID: 23792628. PubMed Central PMCID: PMC3773023. 2. Vouillot, Léna, Aurore Thélie, and Nicolas Pollet...key to address these concerns as CRISPR use grows across the life sciences and to establish standardized...methods. They identify possible off-target sites across the genome and pinpoint the location of mismatches... Digenome Seq 0.1% Cell-free (in vitro) Works across all cell types Must be verified with cell-based...
  9. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives...are a staple in the lab. GFP is used in research across a vast array of biological disciplines and scientists...
  10. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...experimental setup less cumbersome. 3. Portability across multiple systems Terskikh et al. showed that their...
  11. Antibodies 101: Validation

    Type
    Blog Post
    ...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...they require variable expression of your target across samples - it is hard to distinguish between specific...look at RNA expression of your gene of interest across a set of human tissues.  From this data, you see...
  12. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...detection, and control. Int J Pharm. 2018 Oct 25;550(1-2):251-268. doi: 10.1016/j.ijpharm.2018.08.043. Epub...and particle formation. J Pharm Sci. 2015 Feb;104(2):602-11. doi: 10.1002/jps.24259. Epub 2014 Nov 21....as measured by the Edelhoch method. Int J Biol Macromol. 2015;77:260-5. doi: 10.1016/j.ijbiomac.2015.03.027...
  13. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will link biomolecules...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ... of protocols and examples of applications from across disciplines now available. In this post, we’ll ...
  14. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although...cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT expression constructs. (B)...science.abb2920 Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH (2020) CRISPR RNA-guided...
  15. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve...classic rhodamines and their analogs without sacrificing their small size and membrane permeability (Figure...
  16. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ... fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell Sorter (FACs) A machine...photostability of your fluorescent protein when using 2-photon microscopy as this hasn't been well reported...PMID: 18703742. PubMed Central PMCID: PMC2606057. 2. Chtanova, Tatyana, et al. "Dynamics of neutrophil...PMCID: PMC2569002. 3. Egen, Jackson G., et al. "Macrophage and T cell dynamics during the development and...
  17. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    ...MiSeq® run. As shown in Fig. 2, below, differences in input concentration across a 100-fold range (1ng – 100ng...molecules for each sample remain highly normalized across a wide range of input DNA concentrations. This ...normalization of read count and fragment length across samples, NGS data obtained via plexWell™ libraries...
  18. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    ... sets Taking the time to identify and research 2 – 3 potential professions will highlight which skills...professional network. No brainer! Engaging audiences from across the hall, fields, and other professions increases...This short list highlights the ubiquity of skills across professions within and beyond academia. Extracurricular...
  19. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ...axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected...equally important is the way the brain connects across these regions, a process driven by neurons. By ...geography and the way the brain is connected to itself across these regions is important for understanding how...applications; it can enhance therapeutic delivery across broader physical areas of the brain and enable ...
Showing: 121 - 140 of 206 results