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  1. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    ...Scientific Publication, where diverse approaches to peer review were discussed. Science communication opportunities...developmentally programmed regulation of DNA replication and gene transcription during vertebrate embryogenesis...There is a rich and active community of science communication professionals who are thinking about, for example...access affects financial models in information publication. Take a look at the blog ‘Scholarly Kitchen’ ...opportunities If you are interested in the science communications world, you might consider an internship or ... is enjoys broad engagement through science communication with a variety of audiences. She focuses at ... science communication...

  2. What the HEK?

    Type
    Blog Post
    ... This allows them to replicate viral plasmids with the SV40 origin of replication. To generate a virus...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first...cell biology, there are thankfully many other applications that they are useful for! What the HEK is it...cell line and we will highlight some of those applications here.   Virus production HEKs are the go-to ...proteins at high levels. In a case where clinical applications of recombinant proteins are necessary, HEKs ...significantly reduces the cost of production. Research applications There are two areas of research where HEK cells...vulnerable to viral infection. Yes, your cells can catch a cold—or worse! HEK-293T and its derivatives are...

  3. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...protein-only transduction method and its potential applications to CRISPR/Cas9 genome editing.   Protein-only...the HIV protein Tat. Although CPP fusions can translocate a cell membrane, the presence of the CPP may ...that components in the buffer used for protein purification must be helping the untagged protein enter cells...Diego D'Astolfo. CRISPR/Cas9: a potential application of iTOP Having tested iTOP using Cre-lox recombination...showed up to 70% resistance to diphtheria toxin, indicating a 70% frequency of biallelic mutations. Importantly...Importantly, hESCs retained their stemness, indicating that iTOP may be valuable for CRISPR/Cas9 stem cell...potential for off-target effects. Third, protein purification (once a protocol has been established) may be...

  4. 5 Great Apps for Lab Life

    Type
    Blog Post
    ...Gibco organizes its app into four categories: tools, analytics, education and data manager. The tools are...are five FREE science lab apps that have been educational and great for lab life. These apps prevent you...lab notebook. Lab Archives is a multipurpose application that allows you to share your data with colleagues...other nice features. This app also has a great educational section. This section holds the Gibco Cell culture...directions including which buffers to use and that indicates if a sequential digest is needed. The enzyme finder... to this app; the user can search by sequence, category of enzyme, or name to discover all the information... Addgene.org Find Protoctols for Your Research Catch Up On Your Molecular Biology Basics ...

  5. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    ...together a grant application? Are you familiar with the publishing or patent application processes? How ... pages in a Curricula Vitae to showcase your publications, academic performances, and research experiences...information. Think "skills," not just "research and publications” What are the skills you developed through your...and writing was how I got my current job as a Communications Coordinator. Wondering how you can develop ...So You Think You have Skills.   Get Science Communication Practice with a Student Group Like Science in...Blogger! Theresa Liao (@theresaliao) is the Communications Coordinator for the Department of Physics &... a research career, she transitioned into a communications role instead after she completed her MSc in...

  6. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    ...discuss automated protocols. 12:00 PM--Catch up. Finish any communications with my UK team members before they...getting to talk to so many different people! I communicate with customers, software developers, as well ...from interacting with others, and have strong communication skills to tailor your message to any type of... to solve is clear, but the solution is more complicated. You’ll need to be creative to be able to work...team in the UK is five hours ahead, so I’m always catching up in the morning! 9:00 AM--Technical implementation...out. 4:00 PM--Respond to customer help tickets. Catch up on customer help tickets, and respond when I ...technology, software, and science. She loves to advocate for the customer and help build software and technology...

  7. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...ITRs serve as a type of origin of replication to initiate replication of the dsDNA virus. Viral packaging...plasmid while the other plasmids with packaging or replication components are called helper plasmids. Below,... production, they can be helpful for certain applications and aid in ease of use. Our lentiviral guide...AAV are rep and cap. Rep is required for viral replication and also aids in infrequent genome integration...a helper plasmid and mediate efficient viral replication. More information on these viral genes is available...particles. E2A/E2B are responsible for viral replication, with E2B being the primary viral polymerase....is missing the endogenous E1 gene, making it replication deficient. This is typically supplied by the ...

  8. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    ... its function, mimic a biologically relevant modification, or add in fluorescent or chemically reactive...function relationships or, in more practical applications, allow a protein to be more effectively used... you do to any number of experiments with. The catch The rapid expansion of GCE led to the creation of... of many new tools and potential applications. But this focus on development, along with the inherent ...’t give it a try. Optimization Why is GCE so complicated? Well, while the techniques are straightforward...ligation through introducing ncAAs with protein modifications of interest or any of a huge variety of ncAAs...has “10x the impact of PCR. But it’s 100x more complicated.” And Ryan Mehl would know. He’s the director...

  9. How Addgene Creates Content

    Type
    Blog Post
    ...Our educational content  Have you ever wondered how we choose topics for the Addgene blog? Or why we...everything else we do! The goal of Addgene’s educational content Addgene truly believes that sharing speeds...antibody and content team, we identified what applications should be included and what basic information... The guide was then assigned to our science communication specialist, Rachel, to write. Identify expertise...through an editing process and final check before publication.   Do research! Nearly everything we write entails...Guide figures. This requires a fair amount of communication between our content team and our graphic designer...graphic designer upon request that the science communicator "just sketch a western blot out quickly." Final...

  10. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    ...your application a whole week before the deadline. The day of submission, review your application with... their dream.  To help students focus on the application process, and simplify access to academic resources...Scrutinize every opportunity prior to starting an application to assess your eligibility, interest, and career... detailed. Beyond the resume, a cover letter communicates your interest in a specific program, and institution...February represent peak season for internship applications. Thus these noble individuals are inundated ... with a stellar letter of support.        5. Application submission  Set a calendar reminder for every...about outreach among low-income communities to educate parents about resources that can help their children...

  11. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

    Type
    Blog Post
    ...thing that comes to mind. However, ion-based communication is not just for neurons. Non-neuronal cells ...It's just that for non-neural cells, ion-based communication happens at a slightly slower scale.  All living..., and when gap junction-mediated electrical communication is briefly inhibited in the entire worm during...reporting and genetically encoded fluorescent voltage indicators (GEVI) are ideal tools to track bioelectric states...activated proton pump) and ChR2 (blue-light activated cation channel) light-activated channels to hyperpolarize...channelrhodopsin used as a sensitive fluorescent voltage indicator with near-complete desensitization in continuous...extremely exciting emerging field, with many implications for regenerative medicine, evolution of body...

  12. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    ... antibodies may be indicated on occasion, usually after other signal amplification techniques have been...IgG2 or IgG4. This can be important for some applications. Assay use While secondary antibodies are useful..., the flexibility of their use varies across applications. Western blots, for instance, almost always ...For the latter three, secondary antibodies are indicated when the protein of interest is found in low amounts...been ruled out.  Ease of access Signal amplification isn’t the only benefit of secondary antibodies. Finding...quantitative analysis of a sample. Instead, relative quantification can be done using either the direct or the ...antibodies remain the go-to option for signal amplification while providing needed flexibility in assay ...

  13. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ...from the serum. Protein A/G purification Protein A or Protein G purification is one common method. These...reagent that works for your species. Affinity purification Only up to 5% of the IgG antibodies produced...specific for the immunogen. A different type of purification can be performed to isolate antibodies only ...inexpensive. Their short production time and the bulk purification steps keep the cost low compared to other antibody...are particularly useful for certain research applications. Many secondary antibodies are polyclonal, as...target protein thereby greatly increasing signal amplification.  When to stay away from polyclonal antibodies...2018-0065 Fishman JB, Berg EA (2019) Antibody Purification and Storage. Cold Spring Harb Protoc 2019:pdb.top099101...

  14. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ... every genomic locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot induce gene editing...Cas9 ortholog, and the target sequence must be located adjacent to that PAM. To scale up fluorescent CRISPR...this rainbow of colors, Ma et al. performed sophisticated tracking of multiple chromosomal loci in live... do not induce genome editing. If used with catalytically active Cas9, the short, CRISPRainbow gRNA will... PMCID: PMC3918502. Find plasmids from this publication at Addgene. 2. Ma, Hanhui, et al. “Multicolor...PMCID: PMC4364232. Find the plasmids from this publication at Addgene. 3. Ma, Hanhui, et al. “Multiplexed...PMCID: PMC4864854. Find the plasmids from this publication at Addgene. 4. Kiani, Samira, et al. “Cas9 gRNA...

  15. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ... work around this is to "domesticate" your fragment: PCR-based amplification can be used to create silent...and exciting applications, this system is an incredibly powerful tool for cloning complicated constructs...Type IIS sites (such as BsaI or BbsI), that are located on the outside of the cleavage site. As a result...plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning...restriction sites that may not be amenable to "domestication", you might want to consider using an alternative...high-efficiency step. References: Sequential amplification of cloned DNA as tandem multimers using class-IIS...

  16. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...promoter, such as the truncated tyrosine hydroxylase (TH) promoter for catecholamine neurons or glial fibrillary... Flp), to either flip or excise a gene that is located between them. Early recombinase-dependent vector...expression profile at multiple steps from plasmid amplification in bacteria to transgene expression in vivo.... the level of spontaneous recombination. This indicates that the amount of leaky expression can largely...are typically employed during viral plasmid amplification (Fischer, KB. 2019). This means that all DIO...of spontaneous reversion during AAV plasmid amplification. On the left homologous pairs of scrambled cre-recombinase...expression of receptors, post-translational modifications, and other factors that AAV utilizes to gain...

  17. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...mammalian cells often relied on chemical (CaCl2, cationic liposome) or electrical (electric current forming...particles and the virus particles produced could not replicate. However, Mother Nature found a way to re-create...packaging plasmid forming a vector that could create replication competent virus. Even though this system was ...still a potential risk for the generation of replication competent lentiviruses (RCL) especially if you...factors in HIV positive cells could make the virus replication competent. The 3rd generation of lentiviral...transgenes (>8 kb) Potential for generation of replication-competent lentivirus (RCL) Efficient gene transfer...transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human tropism can...

  18. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ... a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium...in living cells and organisms. Unlike chemical indicators, these DNA constructs can be delivered to intact...the sensors discussed below fall into this GECI category.   For calcium sensors, one size doesn...:509-18. Inoue M. Genetically encoded calcium indicators to probe complex brain circuit dynamics in vivo...Kanemaru K, Iino M. Genetically encoded fluorescent indicators for organellar calcium imaging. Biophysical journal...neuronal activity with genetically encoded calcium indicators. Cold Spring Harbor Protocols. 2012 Jun 1;2012...signaling using genetically targetable fluorescent indicators. Nature protocols. 2006 Aug;1(3):1057-65. Resources...

  19. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...host genome, the virus is replication incompetent, meaning it can’t replicate and continue the infection... tools, thought was given on how to keep the replication of virus and its infectivity in check. Engineering...envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein on yet a third plasmid... this safety feature prevents the virus from replicating and spreading in non-target cells, including ....    Viral cargo-specific safety   After replication competency, the second concern with virus safety...diseases in humans, earning this virus a BSL-2 classification. Lentivirus (a type of retrovirus) and gamma...decontaminating liquid waste, remember that the percentage indicates the proportion of the decontaminate in the waste...

  20. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...another method for making targeted modifications. A recent publication by Zhang et al. shows just that -...about a month and a half, without the need for complicated cloning of homology arms. MMEJ: An Introduction...required by MMEJ can easily be added via PCR amplification. Given the inefficiency of HDR for knock-ins... and 3’ junctions, respectively. These results indicate that PITCh is a robust method for genomic insertion...lies in the diversity of nucleases and their applications. It will be interesting to see what new editing...PMID: 26678082. Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015...633. PMID: 26456817. Find plasmids from this publication at Addgene. 3. Zhang, Chi, Xiuhua Meng, Xiaolei...

  21. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...well as suitable applications. Expression System Components of System Application Mammalian expression...CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods.   ...Select the appropriate CRISPR tools for your application: Wildtype Cas9 or Cas9 nickase are appropriate... or introducing mutations and tags, while a catalytically dead dCas9 can be used in conjunction with activator...or more sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair...Cas9-gRNA complex and delivered to cells using cationic lipids. Transient expression of CRISPR components...below: Chemical transfection   Lipid mediated Cationic polymers Calcium phosphate Cells Transformed...

  22. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...up. Applications of photoactivatable fluorescent proteins Some of the most important applications for ...conventional FP imaging. PA-FPs are divided into 2 main categories - those with irreversible and reversible photoactivation...photoactivation - and have enabled sophisticated imaging techniques.     Irreversible photoactivation Irreversible...level of background fluorescence. For any PA-FP application, the brighter the fluorescence the better. More...photoactivation. A high contrast ratio usually indicates that a PA-FP has a low level of spontaneous fluorescence...photostable green and red states. Beyond these imaging applications PA-FPs have been developed for use in yeast,..."Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging." Nature methods...

  23. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...drives and their ramifications. Readers unfamiliar with academia’s challenges with publication and funding ...Connecticut College who has been teaching and communicating science for over two decades, attempts to answer... primarily on fluorescent proteins and their applications, tackles much broader issues in this brief volume...sexism in science, problems with peer-reviewed publications and funding, bioethics in the age of CRISPR,... and her essential-but-unlauded role in the identification of DNA’s double helix. Perhaps to the dismay...dismay of middle-aged scientists, the identification of Green Fluorescent Protein (GFP) and its development...CRISPR of course address some of the ethical implications of using these technologies, but also the rapidity...

  24. An Introduction to Adenovirus

    Type
    Blog Post
    ...and late gene transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the...encapsidate new virions) are not expressed. Note that replication-competent AdV vectors also exist and are used...the final topic, which is that natural AdVs, replication-competent viruses with a recombinant genome, ...Hermiston, T. W. (2003). Multigene expression from a replicating adenovirus using native viral promoters. Mol ...does not require de novo viral gene expression: Implications for gene therapy. Proc Natl Acad Sci U S A, ...in vivo and maintain activated CD8+ T cells: implications for their use as vaccines. Blood, 110(6), 1916...on the Addgene blog Viral Vectors 101: Viral Applications Adenoviral Vector Production and Troubleshooting...

  25. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...helps us to locate interesting addresses in the genome. Let’s now examine how we can locate powerful stretches..., start, and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning of the ...for the genome" techniques Technique What It Locates ChIP-seq Proteins (for our example later, histones...therapeutics. We’ll have more to say about clinical applications of CRISPR in the next section. In order to denote...’s associated histone proteins K4 and K27 – locations of the amino acid lysine in H3 me1, me3, and...aceylation, respectively (these are chemical modifications, or functional groups, found on lysine) Putting...blog biobabel blog Resources on the Addgene Blog Catch Up on Your CRISPR Background with CRISPR 101 Learn...

  26. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...lentiviral plasmids created for a variety of applications including cDNA expression, shRNA-mediated knockdown... looking for some additional information or clarification on these vectors. Read on to find the answers... thus decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells...type or tissues; this is useful in gene therapy applications where a distinct subset of the cellular population...needs to be targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all)...all) Addgene transfer plasmids are replication-deficient meaning they can be used to create virus that is...Which genes are deleted or modified to result in replication deficiency? What percent of the vector components...

  27. Inntags: Innovative Protein Epitope Tagging

    Type
    Blog Post
    ...immunoprecipitation applications, but none of the traditional tags were suitable for both applications. To prove...from non-vertebrate systems. Preliminary testing indicated that inntags IT5, IT6, and IT10 expressed well...and the chaperone Ydj1, both of which display sophisticated interactions with multiple other proteins. Again...detrimental effects on cell size and growth rate, indicating that they compromised protein-protein interactions...These tags are easy to use in multiple common applications. Due to their limited effects on native protein...particularly in single-molecule or high-throughput applications. Perhaps IT5 and IT6 will even replace our older...

  28. Technical Design of a Western Blot

    Type
    Blog Post
    ...original publication and ensure that it includes validation data relevant to your application — and double-check...buffer, consider the subcellular location of your protein. Proteins located in membrane-bound subcompartments...or are looking to preserve specific protein modifications, we recommend doing a little more research into...Blocking solutions Your blocking options come in two categories: protein-based and chemical-based. Before selecting...making membranes inherently “fuzzier” than gels. Quantification If you want to quantify the amounts of protein...available in a pure form, you can perform relative quantification, which measures protein amounts and compares...membrane Increased time Calculate absolute quantification Running a standard curve Uses many wells...

  29. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate natural cellular...editing works in general. Base editors use either catalytically dead Cas9 (dCas9) or a Cas9 nickase (nCas9) ...excision repair (BER), the cellular process that catches mutated U’s in DNA and ideally converts them back...terms gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a matching DNA sequence.   Target strand The ...directed base excision repair proteins. Nature Communications, 12(1), 1384. https://doi.org/10.1038/s41467...double-strand breaks causing deletion, transversion and translocation. Nature Cell Biology, 26(2), 294–304. https:...predictable by machine learning methods. Nature Communications, 12(1), 4902. https://doi.org/10.1038/s41467...

  30. Hot Plasmids - August 2020

    Type
    Blog Post
    ...Fluorescent tools for mammalian cells Nanobody purification CRISPRi and CRISPRa system for hPSCs The CRISPR...CRISPR corner New from the viral service Faster catalysis after yeast-based protein evolution By Joanne ...concept, Sanchez and Ting sought to improve the catalytic efficiency of tobacco etch virus protease (TEV...activity with a fluorescent protein--no affinity purification required! The team used other molecular techniques...evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription factor ...information on where exactly a structure or protein is located within a cell. The Goedhart lab, Gadella lab, and...beads. Workflow of nanobody expression and purification. Iwaki et al., Protein Expr Purif, 2020. ...

  31. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...resources you’ll need ahead of time and mute your notifications during. (Boyes, 2019) If you’ve taken the time... particularly relevant to you. Elect a devil’s advocate. This is the person whose job it is to disagree...think about alternative options. When a devil’s advocate is elected, others may feel they can share opposing...dissenting opinions or ideas. While the best devil’s advocate is one who truly disagrees with the group (because...adamantly for their solution), selecting a devil’s advocate is a good option for any meeting in which the ...other way,” you probably have reason to continue advocating for your idea. Our product team has implemented...also be procrastinators.  If you fall into this category, get out of your head by giving yourself a quick...

  32. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ... than ever to make precise, targeted genome modifications. Cas9 has been modified to enable researchers...instead.  Choosing a Cas9 for site-specific modifications Site-specific deletions and knockouts Some experiments...mutation at a specific site e.g., deleting the catalytic residue of a protein or knocking out a very small...flexibility as to where your sgRNA needs to be located and there may not be a NGG PAM site anywhere to... Beyond SpCas9.” Single base pair edits Is the location and type of edit you need to introduce highly ...Perhaps you need to introduce a single nucleotide modification at a defined amino acid to mimic a patient mutation... mutation or maybe to disrupt a catalytic residue. Good news! You can achieve your precise edit without...

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

    Type
    Blog Post
    ... OriP and induces amplification of the episomes concurrently with DNA amplification in the host cell. ...possibilities for the clinical and therapeutic applications of iPSCs. The generation of human iPSCs (hiPSCs...hiPSCs, reprogramming methods that do not involve modification of the original genome by integrating viral ...where each vector contains a viral origin of DNA replication, OriP and the EBNA-1 sequence that encodes for...are necessary and sufficient for retention and replication of the vectors in human cells [8]. The EBNA-1...cocktail. pCXWB-EBNA-1 lacks oriP and is unable to replicate in human cells, thus expressing additional EBNA...., Recent technological updates and clinical applications of induced pluripotent stem cells. Korean J ...

  34. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ...advantageous to speak multiple languages - that makes communication with customers from around the world easier....their general job duties. Bigger companies have dedicated technical support teams, sometimes comprised of...and email/phone are the most popular ways to communicate with customers. Online live chat is a convenient...instances, variations of the product’s specific application instructions or “optimized steps” can be linked...customers may tell all sorts of details about the application, just not the ones that matter. Therefore, if...the opportunity to develop interpersonal and communication skills. Problem solving and analytical skills...develop an understanding of when to use these communication skills by working with people from many different...

  35. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...Labor’s general guide to the effect on higher education, you can see that those in teaching roles (such...frankly quite surprising. Consortia of higher education institutions submitted letters to the Department...centrally administered, which leads to many complications, including the inability of some institutions...paid. Institutional salary policies are one indicatator of how salaries may changes due to the FLSA. ...of the postdoctoral workforce. Based on our communications with and research on these institutions, we...to the new level:  We are still waiting for an indication of what (most of) Harvard, Johns Hopkins, (most...writing responses to requests for the legal justification of this have not been received. Future actions...

  36. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    ...demonstrate in your application). Beware the myths of scientist “overqualification” but it may not be ...Utilizing existing relationships as part of the job application process can dramatically increase the chances... of a resume can be crucial for a successful application. It takes time to prepare an excellent resume... here for more on timely submission of a job application. 3) Read job descriptions carefully Be open minded...bold. You don’t have to have all of the listed qualifications to apply for a position.  A job description...target someone in a specific department and/or location. The more people you meet the better chance you...

  37. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    ..., just 13 amino acids, but SpyCatcher is a protein. You can fuse SpyCatcher onto things, but even better... Howarth’s lab at the University of Oxford is dedicated to generating new tools to manipulate biology ...particular. They recently introduced the SpyTag/SpyCatcher system, based on a protein isolated from Streptococcus...It came from our work in developing the SpyTag/SpyCatcher system, to be able to grab on to proteins irreversibly... if there are forces acting on them. For the application we were looking at, magnetic isolation of circulating...out the cell capture system.) Cancer and other applications Addgene: What does this technology mean in the...

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

    Type
    Blog Post
    ... the indicated tool. ~ reflects an innovation that may optionally be combined with the indicated tool....genetic variants have been identified in humans and cataloged in the ClinVar database. Previously developed ...prime editing Less constrained by PAM sequence location The prime editor extends the reach of CRISPR genome...editing typically needs to be optimized for each application, and the efficacy varies widely — it still almost...editing variants Four years after their initial publication, the Liu lab returned to the reverse transcriptase...primer binding site. If this tail is degraded, the truncated pegRNA can still guide Cas9 and compete with full...c-Myc nuclear localization sequences Improve translocation to the nucleus ~ ~ ~ ~ ✓ ✓ ✓ ...

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

    Type
    Blog Post
    ...gene at once. Therefore, applications that require simultaneous modification of all alleles of a gene,... recommends designing homology arms with the modification of interest, ordering them from a gene synthesis...with a homologous repair template. It is this application - the creation of sequence-specific knockins ...clones, making it unsuitable for most in vivo applications.  Third, even in the most optimized tissue-culture...tissue culture based approaches that require modification of only a single allele of a gene, such as the...efficiently deliver large repair templates for applications that require large gene fusions. The current...you’ve been able to check out some of the many applications of AAV in some of our other blog posts and have...

  40. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...knowing that where you see fluorescence indicates your target’s location. Some other terms you might see associated...immunofluorescence? Generally speaking, IF is an application in which antibodies that have been joined to ...bind to a single primary, resulting in signal amplification. Another reason is cost effectiveness. Generating...distribute antibodies validate them in a variety of applications, including IF (or IHC or ICC), and will share...will share their findings either through their publications or as a review on a company website. It is important...Resources on the Addgene blog Learn about research applications of antibodies in Introduction to Antibodies ... for Me? Read about another common antibody application in The Basics of Western Blotting Resources ...
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