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We narrowed to 770 results for: VARS

Showing: 141 - 160 of 770 results

  1. Antibodies 101: Isotypes

    Type
    Blog Post
    ...downstream of the variable region gene in this order: μ, δ, γ, ε, and α. An antibody variable gene is initially...heavy chain, light chain, constant region, and variable region.   Why are antibody isotypes important... heavy chain constant domain it pairs with its variable chain domain. B cells start by co-expressing IgM...antibodies, but later in an immune response, the variable region of an antibody is paired with any of the...intervening DNA sequence is deleted, which places the variable region next to a different constant domain gene...heavy-chain constant domain genes, placing the variable domain genes next to a different constant domain...the B cell expressing an antibody with the same variable domain but a different heavy-chain constant domain...

  2. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...’ll dive into the opposite: AAV serotypes with varying tissue tropism, allowing them to target particular...shell packages the vector’s genetic material, and variations between serotypes can change how the viral particle...be present on different cell types and sometimes vary between target species or even strains. Pro tip!...serotypes in this post. However, tissue tropism varies both between species and even between strains of...currently understand it. Tropism depends on many variables, so be sure to check the literature for previous...applications. Cross-reactivity of AAV serotypes varies by species and route of administration (Wu, Asokan...efficiency in pancreatic beta and alpha cells utilizing various AAV serotypes. Scientific Reports, 15(1), 20927...

  3. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    ...very soon. Not only is CRISPR having effects on various biological disciplines, the base technology itself...itself is constantly improving. Cas9 variants have been modified for genome editing, activating gene expression...specificity of the Cas9 nuclease with engineered variants: eSpCas9, SpCas9-HF1, & HypaCas9. The off-target...off-target effects” have included using a Cas9 nickase variant, lowering Cas9 expression, and truncating gRNA ...Zhang lab at the Broad Institute, the Joung lab at Harvard Medical School, and the Doudna Lab at Berkeley ...of the HNH/RuvC groove, Slaymaker et al. made a variety of alanine substitutions throughout the groove ...off-target cuts when compared to WT SpCas9 across a variety of genomic sites. Knowledge of Cas9’s dynamic structure...

  4. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...is supplied on its own expression plasmid or in various combinations, while the cognate att sites are supplied... BRET ratio), which allows for compensation of varying fusion protein expression levels. Zhang et al...ensures that all screened plasmids contain a gene variant* and is amenable to both high and low throughput...protein with cell specific expression containing a variety of N- and C- terminal tags. The Santalucía Lab ...specifically express your new chimeric protein in a variety of model systems. The MultiSite Gateway Kit contains...constructed with the MuLE toolbox have been used for a variety of experimental purposes including (1) single or...been used to study transport characteristics of various Golgi and plasma membrane reporters as well as ...

  5. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...exciting year for genome engineering research. A variety of new tools came out including the single base...base editors, Casilio, CombiGEM, and a variety of pooled libraries. Not all of these technologies were without... use to express both SpCas9 and your gRNA in a variety of mammalian cell types. Purpose  SpCas9 This plasmid...CAG promoter that the Zhang lab has tested in a variety of mammalian cell lines. Cas9, of course, is the...particular backbone can be used to transfect a variety of mammalian cells and can be modified for viral...express both SpCas9 and your gRNA of interest in a variety of mammalian cell lines. Alternative forms of this...GFP-tagged and un-tagged versions of SpCas9 and further variants replace SpCas9 with SpCas9n (nickase), which nicks...

  6. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...MIT, Alvin Hsu Harvard University, and David R. Liu from the Broad Institute and Harvard University. Over...MIT, Alvin Hsu Harvard University, and David R. Liu from the Broad Institute and Harvard University. Max...sophisticated analyses, we first visualized our data in various ways. Smoothing our plasmid order data with a 7...parameter. We approximated the data likelihood with multivariate Gauss-Hermite quadrature, since the analytical...intractable integral of the product of a Poisson and multivariate log-normal likelihoods. Effects of weekends...applications.       Alvin Hsu is a graduate student at Harvard University. He is interested in using selection...Professor of Chemistry and Chemical Biology at Harvard University; and Howard Hughes Medical Institute...

  7. Typing CRISPR Systems

    Type
    Blog Post
    ... simple hierarchy: class to type to subtype to variant. Despite the seemingly complicated steps to arrive...on main function and composition; subtypes and variants are typically based on species of origin. There...currently two classes, six types, 33 subtypes, and 17 variants. Figure 1: Overview of the CRISPR classification...hierarchy flows from Class to type to subtype to variant (shown in italics). Created with BioRender.com....machinery from other CRISPR systems to carry out various functions. Class 2 Class 2 CRISPR systems rely ...others have become more notable in recent years. A variant of V-U, paradoxically called V-K, is a CRISPR-associated...that can insert large fragments of DNA. The V-F variants, which are among the smallest Cas enzymes (400...

  8. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...measure the frequency of each variant during the selection process. Each variant is assigned a functional ...of a plasmid library that expresses all desired variants of a protein. Applying a selective pressure winnows...winnows the pool down to plasmids expressing variants with optimal function. High-throughput DNA sequencing...mutational scanning Hundreds of thousands of protein variants can be tested in one experiment. There are many...experiments, libraries of DNA fragments that contain variable bases (NNN) at any given codon site of interest...opposite wild-type DNA strand is nicked by the BbvCI variant not used in step 1. Then this strand is degraded... and cost. You need at least 100-200 reads per variant, so the more codons you test in one library, the...

  9. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ... released a new variant of jGCaMP7 (keep reading) and will be releasing more variants in the next few ...few months (see “coming soon”). The 7b variant (pGP-AAV-syn-FLEX-jGCaMP7b-WPRE) exhibits the brightest resting...processes like dendrites and axons. Find the 7b variant of jGCaMP7 in AAV1 at Addgene! See all calcium ...membrane localization and brightness. The F102G variant (pAAV.hSyn-FLEX.iGABASnFR.F102G) is noticeably ...responses to GABA. Find the iGABASnFR and its F102G variant in AAV1 at Addgene! Archon1, a voltage sensor AAV...neurons. 105321  AAV1  pGP-AAV-syn-jGCaMP7c variant 1513-WPRE 7c exhibits high contrast between peak...reduced. 105322  AAV1  pGP-AAV-syn-FLEX-jGCaMP7c variant 1513-WPRE7c exhibits high contrast between peak...

  10. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...pathogenic genetic variants have been identified in humans and cataloged in the ClinVar database. Previously... potential to correct only a minority of those variants in most cell types. But prime editing, a CRISPR...innovations by the Liu lab and others have led to a variety of prime editing tools. We’ll summarize the major...optimized for each application, and the efficacy varies widely — it still almost always results in higher...research, the Liu lab created and evaluated 19 PE1 variants with RT mutations known to increase activity, ...Even in PE3 and PE3b, prime editing efficiency can vary widely — so its inventors kept optimizing it. Using...PE2max, PE3max, etc. PE6: Specialized prime editing variants Four years after their initial publication, the...

  11. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    ...-AAV-syn-jGCaMP7c variant 1513-WPRE (105321-AAV1) pGP-AAV-syn-FLEX-jGCaMP7c variant 1513-WPRE (105322-...ul) samples of EGFP-expressing AAV packaged in various serotypes. This collection is now complete! pAAV-CAG-GFP...vectors to more serotypes. We currently package four variants of these biosensors, which are optimized for specific...relative to baseline fluorescence, as the GCaMP7f variant, and is approximately 5-fold brighter than GCaMP6s...GCaMP6s. jGCaMP7f is the fastest responding variant. jGCaMP7b exhibits the brightest resting fluorescence...fluorescence from inactive neurons is reduced. This variant has an approximately 12-fold increase in brightness.... We recently released several recombinases in various serotypes, and will continue to grow this collection...

  12. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    ...the most recent advances and how some of these variants may be appropriate for your specific research ...But Cas9s found in other species can recognize a variety of different PAM sequences. Researchers have also...isolated from different organisms and engineered Cas9 variants that were able to target different PAM sites (... be targeted (Hu et al., 2018).  These altered variants can allow for targeting of your genomic loci of...off-site activity of Sniper-Cas9 to different Cas9 variants using gRNAs of different lengths (Lee et al., ...continue to characterize CRISPR nucleases from a variety of sources, it is likely that the toolkit available...genome, there are more and more Cas enzymes and variants to choose from. While no enzyme is perfect for...

  13. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...different plasmids. Harvard’s DRSC/TRiP Functional Genomics Resource Center has various plasmids for overexpression...blogger Jon Chow, an immunology PhD student at Harvard University. In my previous two posts, I’ve described...Random insertion has the advantage that you can get varying expression levels of your transgene in different...comparisons between lines expressing different variants of YFG because the transgenes are found in the...that are anywhere from light yellow to red. The variation in color depends on how transcriptionally active...interest, such as the intestines, can be dissected at various times throughout the lifespan of the fly to determine...Chow. Jon Chow is finishing his immunology PhD at Harvard University. He has had nearly a decade of experience...

  14. Deep Dive: qPCR

    Type
    Blog Post
    ...Yes Agarose gel Table 1: Features of various qPCR chemistries. Adapted from Wong, 2018   ...should be 60-65°C For qPCR, the important primer variable is the annealing temperature, not the melting ...reference genes have shown to have as much as sixfold variation in expression levels reported (Biassoni, 2014)...negative error on the other will introduce unwanted variation into your data. Try different types of pipettes... within each set of triplicates to understand variation due to technical error.  Plate readouts can be...type or tubes, sealing method, and other small variations in consumables. Track and record these details...Reference Dyes Whatever your pipetting method, some variation will occur. A passive reference dye, such as ROX...

  15. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ... an optogenetics experiment work? What are the various tools and components necessary to run it? And, ...any optogenetic experiment. There are numerous variants for the different types of opsins available and...constructs available through the vector cores of various universities as well as Addgene. See the Deisseroth...cell-type specificity, the kinetics of the opsin variant, and other particularities that best suit your ...these reviews for a discussion of the different variants [1, 2, 4]. And, of course, the best way to learn...learn about the many different opsin variants and how they can be used is to consult the many primary research...has developed many of these AAV constructs with various promoters and other genetic manipulations to help...

  16. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ... Germline variation contributes to false negatives in CRISPR-based experiments with varying burden across...Beroukhim labs at the Broad Institute of MIT and Harvard, Dana-Farber Cancer Institute, and Koch Institute...the effects of ancestry and germline sequence variation in cancer cell lines took them down unexpected... screens based on the cells’ germline genetic variations. They then wanted to identify cancer-associated...correct, either. However, when they analyzed the variations in gene expression, they found that for approximately...genome does not represent the diversity of genetic variation across the human population. Rather, it primarily...designed that target regions with high genomic variability.   Moving forward It took a five-team response...

  17. Science Communication Snapshot: DayCon 2015

    Type
    Blog Post
    ...advancing and sharing science! In association with the Harvard graduate student organization Science in the News...public that provides an accessible introduction to various scientific topics. Over twenty graduate student... with the public. The organization runs a wide variety of programs, including fall and spring lecture ...and Reddit AMAs. Their website also provides a variety of short- and long-form written content about timely...co-director Steph Guerra, a 3rd year graduate student at Harvard, who wanted to provide a conference-style experience...and content level. Sponsorship from Addgene and Harvard Integrated Life Sciences (HILS) made it possible... the biomedical sciences and decided to offer a varied format event to keep the audience engaged. The ...

  18. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...CRISPR and optogenetic techniques to develop a variety of photoactivatable CRISPR tools. These tools allow...genome editing process. Read on to learn about the various light-controlled CRISPR tools available to researchers...activate gene transcription. The researchers tested a variety of combinations to optimize both fusion proteins...photoactivatable repressor (dCas9). The activity of all variants was reversible and repeatable. Since these initial...CRISPR–Cas9 activity switching via a novel optogenetic variant of AcrIIA4 (CASANOVA). As shown in the figure below...this article). The group first tested photocaging various lysines in Cas9 to determine which best deactivated...Publication System nickname Photoactivatable moiety Cas9 variatn CRISPR protein application Split Systems / Light-induced...

  19. Hot Plasmids - February 2022

    Type
    Blog Post
    ...Here, Liu et al. engineer a chimeric single chain variable fragment (scFv), grafting the binding region of...improve its stability in cells, and use it in a variety of applications. This set of new anti-FLAG frankebodies... (c) Applications of anti-FLAG-FB-GFP labeling various targets with nuclear (top, 4x-FLAG-mCherry-H2B)... Anti-FLAG and anti-HA frankenbodies have a variety of applications, including live-cell imaging, single-molecule...optimizing enzyme activity using an enhanced Cas12a variant (enAsCas12a), minimizing off-target effects by ... reducing recombination by finding acceptable variations in DR sequences. Figure 2: comparison ... Various Addgenies...

  20. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    ... even distinguish between protein variants when used with a variant-specific antibody. TRIM21, an E3 ubiquitin...inactive cas9 (e.g. dCas9) has been fused to a variety of protein modules to repress or activate gene ...complexes to any genomic locus in any cell type. By varying which sgRNAs are used with FIRE-Cas9, experiments...then induced cultures of these transformants with various concentrations of estradiol and progesterone and...transcription factors to control the level of random variation, or noise, in reporter expression. To do so, one... Various Addgenies...
Showing: 141 - 160 of 770 results