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We narrowed to 932 results for: COMP

Showing: 261 - 280 of 932 results

  1. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...when studying them since their complete knockdown is lethal. Incomplete or titratable knockdown, however...generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters et al., 2019 with permission... Top: A Tn7 transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes...ability to synthesize diaminopimelic acid (DAP), a component of the cell wall. Once in the recipient cell, ...Bottom: An ICE element containing the CRISPRi components is transferred to recipient bacteria by bi-parental...was variable. They measured transfer rates by comparing the number of colonies that grew on a selective... gRNA were more sensitive to trimethoprim when compared to control gRNA treated or untreated bacteria....

  2. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...realized how perfectly flat the complex is on this surface”….where the complex is docked on the membrane of...using 3D printing technology. The tagline for our company is “…where molecules become real”.  We sometimes... the real thing. Nevertheless, the models are compelling, and I encourage you to hold one. Why do we ... They then start talking to themselves as they compare the features displayed on the model with their ... close to its reported cleavage site on the complementary DNA strand. And this is why we love working ... that was not “poised for cleavage”. Jacob computationally added the non-template strand to the Jinek ...double-stranded DNA, recognize a PAM sequence, allow the complementary base pairing between the guide RNA and the target...

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

    Type
    Blog Post
    ... millions of people die from cancer-related complications each year. Despite these grim statistics, new...cancer, really? While cancer is extraordinarily complex at the molecular level, at the cellular level there...keeping cells from dividing out of control. One may compare the roles of proto-oncogenes and tumor suppressors...cancer research has focused on picking apart the complex cellular pathways involved in the development of...for this elusive target. Recent improvements in computer modeling have enabled researchers to begin discovering... prevalent in human cancers, and synthesizing compounds to bind specifically to their mutated residues...biggest challenges facing researchers today is the complexity of the pathways involved in the development of...

  4. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...antibody-conjugated labeling. Even though the two compounds are completely different in nature (eGFP being a protein...results in a higher-wavelength emission spectrum compared to APC. This happens due to a process called Fluorescence... red laser excites at 640 nm (Table 1), while comparing the two fluorophore's excitation spectra around...think of brightness levels on your smartphone or computer screen), and you should be able to properly label...the same color panel. In this case, make sure to compare the excitation/emission spectra of your reporter...whether and to what extent they overlap. Figure 4 compares the spectra of enhanced GFP (eGFP) and FITC, the...Make sure to check out our blog post on color compensation to learn more about them. Alright, let's build...

  5. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...referred to as “complex retroviruses ” (Cullen, 1992) because of their more complicated genomes. Here, ...biology. “Retroviruses” is an umbrella term which encompasses 1.) the gamma-retroviruses — that have been developed...whole family; however, you can use Figure 1 to compare the genomes of these two family members.   ...beginning to realize that the situation is already complex enough that a deep understanding of retroviral ...Because it is made at the same time as the other components of the vector particle, some of the RT protein...In both cases, the lipid membrane is the only component that is not dependent on viral genes. It is derived...Mechanism of action of regulatory proteins encoded by complex retroviruses. Microbiological Reviews, 56(3), 375...

  6. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    ... if you can begin simply and build up to more complexity if needed. And since we live in the digital age... biology and ecology can make for beautifully compelling photos and videos, while molecular, micro, and.... Can you convey enough information to tell a compelling story in an image or do you need a full exhibit...fair, or a semester-long after school program? Comparing your desired outcome to the limitations and potential...expertise needed can increase exponentially with complexity. Podcasts Medium Easy for audience to engage... needed. Traditional media formats are quite competitive while digital can be oversaturated. Transition...Photography Medium (high for microscopy) Visually compelling. Easy to convey a large number of details without...

  7. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...of the vector components are HIV-based? A7: No single plasmid contains all the components necessary to ...The components are divided as follows: Transfer vectors contain minimal cis-acting HIV components: LTRs...decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells.  Q4: How...targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all) Addgene transfer...LTRs, PPT, RRE, and psi packaging signal. Viral components typically total <1.5kb, which is rarely more ...are: 1) the potential generation of replication competent virus [usually HIV-1]; and 2) the potential for...

  8. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...the cell must have enough nutrients available to complete the entire round of replication. Initiation is...initiate replication. But pSC101 requires a third component: RepA, a protein encoded by pSC101 itself. RepA... are efficient for the host, as no additional components are necessary. Iterons are not the only tool ...Inc, also encoded by the ColIb-P9 plasmid, is complementary to part of the sequence forming this pseudoknot...restraining further replication.   Figure 3: Comparison of replication regulation mechanisms of stringently...replication. At high copy numbers, RepA proteins form complexes between iterons on different plasmids in a process...propagation by limiting their copy number and compensating for the additional metabolic cost imposed on...

  9. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...have the full picture of the puzzle, the QC team compares it to the plasmid information or depositor-provided...depositor-provided sequences to confirm the correct plasmid components. For more information on this process, see our...small sample sizes, small regions of interest, or complex sequences such as G-C rich areas, secondary structures... NGS technology, due to the high-throughput compatibility. However, there are some instances where we ...thousands or even millions of base pairs. Several companies have worked on this technology, but Oxford Nanopore...sequence an entire plasmid in one read. Runs can be completed in anywhere from under an hour to 72 hours, with...data output. This method is great for resolving complex and repetitive sequences, but can be costly, low-throughput...

  10. Hot Plasmids - November 2023

    Type
    Blog Post
    ...  Here's what you'll find in this post:  New compact, efficient prime editors A versatile and precise...tape Teaching an old antibody new tricks   New compact, efficient prime editors By Susanna Stroik The ....), while PE6c and PE6d were most efficient at complex edits, such as long or highly structured templates...Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Cell, 186(18), 3983–4002...interchangeable set of fusogenic and targeting components to reprogram delivery vehicles like lentiviral...DIRECTED particles where the fusion and targeting components are interchangeable. Image from Strebinger et...the range of targetable cell types. DIRECTED is compatible with fusogens from different viral families and...

  11. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...intense amount of competition between microorganisms for available resources. This competition drives microbes...antibiotic compounds. In such a case, the microbe itself would be unharmed by the compound it produces...treat infections, we are treating microbes with compounds similar to those they have been battling for centuries...treatment regiment might wipe out bacteria that are completely susceptible to a drug while a full course of ...likely to develop over time. It isn’t feasible to completely eliminate the development of resistance; it’s...hygiene standards. Handwashing is an essential component of preventing infection, as is cleaning and disinfecting...

  12. From our Table to Yours: An Inside Look at Lunch at Addgene

    Type
    Blog Post
    ...lab. Iron Chef-style competitions Every few months, we’ll have a cooking competition featuring a selected... eat together every day, sharing ideas for the company, but also talking about sports, movies, and just..., which is either an indication of the stiff competition or says something about my cooking.   This is... to organize. We’ll dedicate a post to these competitions in the future, and hopefully include some of...To-Do list long enough that there is no hope of accomplishing it all. How do we encourage people to take a... Hack Days, and we’ve seen great success at accomplishing “we-should-really-make-time-for-that” type projects...

  13. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...likely not a coincidence that the AAV genome is composed of single stranded DNA, which is particularly ... interest, ordering them from a gene synthesis company such as IDT or Genscript, and cloning them sequentially...infection with adeno-cre. AAV-based gene editing compared to CRISPR editing CRISPR, of course, has started...and virtually never exceeds 40%. While this is comparable or better than the efficiency of most current...systems. Ohmori et al 2017 use a dual vector system composed of SaCas9 and a gRNA in one AAV and a repair template...researchers have devised methods to deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied...2017 recently showed that by delivering CRISPR components as RNPs and delivering two halves of a repair...

  14. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...based on its library frequency before selection compared to its library frequency after selection. Key ...and easy technique that can be used to generate complex libraries of mutant plasmids ready for deep mutational...libraries, they all have inherent shortcomings compared to one pot saturation mutagenesis. Error-prone...vector with Gibson assembly. While this is a comprehensive mutagenesis method, a drawback to using Gibson...scanning libraries. PFunkel: Of prior existing comprehensive mutagenesis methods, PFunkel offers the best...second universal primer is used to synthesize a complementary mutant strand (step 2 and 3). Then the uracil-labeled...PCR-based approach for generating a customizable comprehensive mutagenesis library that’s ready to be tested...

  15. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...now available. However, it can be difficult to compare their performance, as most translation initiation...directly engages the ribosome for translational complex assembly. PhD student Dejan Gagoski used SITS ...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of proteins...-tagged human ORF clones to allow testing and comparison of different cell-free expression systems; the... construct barcoded, locus-specific libraries compatible with Illumina next generation sequencing in order...libraries using a 2-step PCR-based protocol and are compatible with single-end or paired-end read flow cells...21408-21413. PUF Assembly Kit PUF proteins are composed of repeat segments that can be engineered to bind...

  16. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ... Now that you’ve completed a resource budget, and know how to decide which components are the most critical...an account of the material resources needed to accomplish your science, and can include: equipment, reagents...then start identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline...research success. If you can get away with using computer modeling and collaborations rather than using ... most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE...equipment. That’s why a resource budget can help you compare the monetary and empirical value that helps you...from academia altogether and instead started a company around his own research. He was clear on his science...

  17. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...Using AAV vectors in complex biological systems can be tricky at best, and downright infuriating at worst...Serotypes Serotype refers to the different protein compositions of AAV capsids, which determine how a virus ...what works great in one system or cell type can completely fail in a different system (Issa et al., 2023...designing and testing your AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9 in two ...Taschenberger et al., 2017).     Figure 3: Comparison of two astrocyte selective promoters demonstrating...Konishi, Y., Hosokawa, Y., & Konishi, H. (2014). A comparative analysis of constitutive promoters located in...vector-mediated transduction of the corticospinal tract: Comparison of four promoters. Gene Therapy, 28(1–2), 56–...

  18. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...reduced transformation and transfection efficiency compared to the monomeric version of the plasmid (Maucksch...multimers have additional origins of replication compared to monomers, they are also able to replicate at...bacterial cells with multimers grow more slowly compared to those containing the monomeric version of the...and more prone to being lost in bacterial cells compared to monomers, although this can be prevented via...cannot be used to detect plasmid multimers. Some companies provide NGS whole plasmid sequencing services ...frequency of multimerization, and in some cases completely prevent the formation of plasmid multimers (Bedbrook...produced the highest yield of monomeric plasmid compared to DH5α. However, more research is needed to determine...

  19. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...binary systems. In recent years, a number of more complex systems have also been developed which don’t fit... Bujard, both the promoter and transactivator components of the system have been engineered and improved...revolutionized genetic studies in flies. An additional component, the GAL80 repressor, adds a repressible element...UAS-driven gene expression. To further increase the complexity of the system, researchers have used split GAL4... developed by Lai and Lee. LexA/lexAop is a complementary system to GAL4/UAS that functions in essentially... cells and ~24,000-fold in human HeLa cells as compared to basal expression. Co-expression of repressor...because it allows them to be used together for complex genetic analysis in Drosophila or for precise control...

  20. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...to detect a single, specific protein within the complex mix derived from cells or tissues. Westerns are...sample per lane, onto the top of a resolving gel composed of the crosslinked polymer acrylamide. In addition...proteins are immobilized onto a membrane, usually composed of a nitrocellulose or polyvinylidene difluoride...membrane.  In a wet transfer system the sandwich is completely submerged in a tank filled with a transfer buffer...confirming that the protein is the expected size as compared to the protein standards.  Protein abundance In...loading control when assessing protein abundance or comparing samples across lanes.  For a typical loading control...present in all samples. Uneven sample loading or incomplete protein transfer causes inconsistent levels of...
Showing: 261 - 280 of 932 results