...to dramatically increase the number of CRISPR applications. The first non-Cas9 CRISPR protein adapted for...doi.org/10.1038/nbt.3609 Find plasmids from this publication at Addgene. Kim E, Koo T, Park SW, Kim D, Kim...10.1038/ncomms14500 Find plasmids from this publication at Addgene. Kleinstiver BP, Prew MS, Tsai SQ...10.1038/nature14592 Find plasmids from this publication at Addgene. Kleinstiver BP, Tsai SQ, Prew MS...doi.org/10.1038/nbt.3620 Find plasmids from this publication at Addgene. Hu JH, Miller SM, Geurts MH, Tang...10.1038/nature14299 Find plasmids from this publication at Addgene. Walton RT, Christie KA, Whittaker.../j.cell.2015.09.038 Find plasmids from this publication at Addgene. Additional Resources on the Addgene...

...activity of CcdB. Applications of ccdB in plasmid cloning Perhaps the most widely used application of CcdB is...used as biological tools for a wide range of applications in molecular biology or in vivo studies. Let...types of plasmid addiction systems and their applications. The discovery of toxin-antitoxin systems Toxin-antitoxin...CcdA (antitoxin) where CcdB inferes with DNA replication by binding the GyrA subunit of DNA gyrase and...construct. Other toxin-antitoxin systems and their applications MazE/MazF Aside from CcdB/CcdA, there are many...Kulakauskas, Saulius, Arvydas Lubys, and S. Dusko Ehrlich. "DNA restriction-modification systems mediate ...

...performs well in many different antibody-based applications.  IPI’s recombinant version is a fusion of mouse... reused from Anderson et al., 2024 under CC BY license.    Find Anti-V5 [IPI-SV5-Pk1] Antibody here...adapted from Kroupova et al., 2024 under CC BY license. This construct can be easily expressed and...Image reused from Cao et al., 2024 under CC-BY license.  These optimized fluorescent proteins will...cellular processes in C. elegans: mScarlet3 for applications needing intense brightness (such as low-expression...first use of CRISPR/Cas9 in M. abscessus. The simplicity and efficiency of this dual-plasmid approach ...Image reused from Neo et al., 2024 under CC-BY license.   Find plasmids for CRISPR/Cas9 in M. abscessus...

... citations for recent articles. Of course, a publication in Science or Nature is always nice, but in the... lesser-known journal as it is in a top-tier publication. Accessibility is surely also behind the increase...increase in citations for older publications. (As an interesting aside, Nature recently reported that the...in a solution.) But are citations and journal publications really the best way to measure impact? As that...and what it’s made of. Increasingly, journal publications alone aren’t enough. Scientists are sharing ...consider and quantify the impact of scientific publications. Ventures like Impact Story seek to find ways...

...decades. Widely used across various research applications, they have become indispensable tools for manipulating...-lasting changes in cells. However, for some applications, integration might be considered a double-edged... is distributed across multiple plasmids. For simplicity, we’ll use the second-generation system as an...dividing cells because they lack an origin of replication (Banasik and McCray, 2010). However, IDLV genomes... certain types of clinical or basic research applications. Considerations for IDLV use If you’re reading...Integrase-defective lentiviral vectors: progress and applications. Gene Therapy, 17(2), 150–157. https://doi.org...lentiviral vector: prospects for safe clinical applications. PeerJ, 10, e13704. https://doi.org/10.7717/...

...current CRISPR research focuses on its potential applications for human medicine (Waltz, 2016), the potential...selected the correct CRISPR components for your application, it is time to deliver these components to your...detergent, Polyethylene Glycol (PEG), although the application of this approach is limited to protoplast cells...find many plasmids for a wide variety of CRISPR applications in plants available through Addgene. In addition..., we highly recommend reading the associated publications or protocols to get the most out of the plasmid...N (2016) The CRISPR/Cas Genome-Editing Tool: Application in Improvement of Crops. Front Plant Sci 7: ....10.1073/pnas.1818290116 Zhu H, Li C, Gao C (2020) Applications of CRISPR–Cas in agriculture and plant biotechnology...

... February, the Office of Science and Technology Policy issued a memorandum calling on Federal agencies...in annual R&D expenditures to support increased public access to the results of federally-funded research...an organization called Free the Data launched a public database to gather the results of Myriad's genetic...support for open access to peer-reviewed journal publications, although many don’t necessarily publish in ...gave preference to journal access granted via a public repository or authors’ own web sites. 9)      The...to share science. Image credit: Judy Breck, via flickr  ...

... ReFigure. Reading and exploration including replications and experiments resulting in “negative data”...small selection of data found in full-length publications. As early career researchers, we wondered how...performance as in this antibody ReFigure Analyzing replications as in this ReFigure about whether Natronobacterium...reagent performs well, or you might publish a replication experiment that you conducted as part of setting...papers as they are not included in traditional publications. By submitting your data to a repository (Figshare...-registration, preprints, data, software, micropublications) in grant proposals. ReFigures are one such...

...Prepare by going sleuthing on the lab website, publication records, LinkedIn, ResearchGate or Twitter. Be...the management style? How does the PI deal with conflicts in the lab? Do they motivate with the appropriate...the appropriate first authorship position on a publication of her project and made another lab member first...professional status – including belittling opinions, public professional humiliation, accusations regarding... Is the work (topic and/or technical aspects) applicable to industry if this path interests you  (e.g....connections such as serving on Boards? Does the publication record show evidence of industry collaboration...out PI intervenes if there are interpersonal conflicts PI has no idea who is dating whom Expanding...

...Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS...tet-systems-legacy-products/tet-on-and-tet-off-2nd-generation Mullick, A., Xu, Y., Warren, R. et al. The cumate gene-... PMCID: PMC2942869. Find plasmids from this publication at Addgene. del Valle Rodriguez, Alberto, Dominic... PMCID: PMC2883883. Find plasmids from this publication at Addgene. Wei, Xing, Christopher J. Potter... PMCID: PMC3846601. Find plasmids from this publication at Addgene. Riabinina, et al. “Improved and ... PMCID: PMC4344399. Find plasmids from this publication at Addgene. Riabinina O, Potter CJ. The Q-System...

...athletes, as well as the ethical and health policy implications arising from sport-related concussion.   ...are sincerely interested in connecting with the public in a meaningful way, and not solely to ensure a...doing so—can prove invaluable in the pursuit of public engagement. Studying science communication is ... research topic on concussion in sport and the public’s perception of head injury risk. In completing ...of unsettled science. Understanding just how the public utilizes insights gleaned from scientific research...

...interested public.” Subscribers to Scientific America would be considered the interested public and it would...Image courtesy of Ron Mader, via Flickr, under the Creative Commons license     ...podcasts A series of Tweets breaking down a new publication that could advance treatments for a rare disease...emails, most science communication work does not explicitly exclude people outside of the original recipients...

...orthologs similar to TtAgo and PfAgo, another thermophilic Argonaute. They identified NgAgo and showed ...template is supplied. Find Plasmids from the NgAgo Publication at Addgene Cas9 and NgAgo head to head NgAgo ...differences in cleavage efficiency. If this finding is replicated in larger studies, it will represent a key advantage...site specified by the gDNA. Depending on your application, this could be favorable or unfavorable. In a...PubMed PMID: 27136078. Find plasmids from this publication at Addgene. 2. Swarts, Daan C., et al.  “DNA-... PMCID: PMC4697943. Find plasmids from this publication at Addgene. 3. Swarts, Daan C., et al. “Argonaute...

...experienced. Finding an antibody that works for your application, specifically detects your protein, is species...and have been widely used across species and applications. In this blog, we will review the most common...not exist, or they don’t exist for a specific application, or they exist but are unreliable for a variety...almost certainly already been optimized for your application. Depending on the tag, directly imaging or tracking...our affinity purification tag blog!).   Applications of epitope tags   The tags A number of...with moderate to high success rates across all applications. You can find FLAG antibodies on Addgene’s catalog...

...need? Multiple first or corresponding author publications (published, not in preparation) A history of...The other items will follow. If you get more publications, you will be more competitive for fellowships... project. Furthermore, Group Leader position applications require 3 letters of reference- the graduate...spurious yet intriguing findings that fail to replicate. Plan experiments logically and strategically ...and exploration Researchers face a fundamental conflict. Science is about exploration, discovering new...informal recommendations, more positive grant application and manuscript reviews, etc. You'll also meet...teach classes. Therefore, seek out management and conflict resolution workshops. Get teaching experience ...

...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... incubate your cells with virus at different multiplicity of infection (MOI=ratio of virus to cells). ...

...regions flanking the target site).  The best applications for dsDNA inserts include: large insertions ...targeted, it’s best to determine the direction of replication through your region of interest and design an...it’s easier to not determine the direction of replication and instead design oligos that target both strands...correct DNA mismatches that occur during DNA replication. There are two ways to avoid activating MMR: ...Uses in Recombineering Substrate Best Suited Application Recombination Frequency Special Considerations...from a plasmid with a heat sensitive origin of replication.  Once the recombineering system is no longer...defective prophage is to use mini-λ, a defective non-replicating, circular piece of phage DNA, that when introduced...

...or DNA base pair mismatches (8-11). For some applications, there cannot be any end-joining activity, and...regions. Why use a high fidelity ligase? Numerous applications have been developed to take advantage of the...Taq and other thermostable DNA ligases. These applications detect specific nucleotide sequences with high...Ligase are not replacements for T4 DNA Ligase in applications such as routine cloning or DNA library preparation...al. "Ligase chain reaction (LCR)-overview and applications." PCR Methods Appl 3.4 (1994): S51-64. PubMed... detection system for direct identification of allelic composition of mutated DNA in a mixed population...to accelerate characterization of nucleic acid metabolic enzymes." Nucleic acids research 44.2 (2015): ...

...tropism, replication kinetics, and cell damage profiling of SARS-CoV-2 and SARS-CoV with implications for ... Another example of reagents that have broad applications but have proven useful to the study of SARS-...vectors VSV-G and psPAX2, which were used in a publication by Xiuyuan Ou, et al. that characterized the ... requested these plasmids from Addgene. This publication then resulted in the development of a new plasmid...scientific community. Deposits of new reagents, policies from journals and granting agencies requiring ...

...objects that exist on the internet so that when you click on the link it will take you directly to the original...item. Given a specific Addgene ID # listed in a publication, you can quickly pull up the associated plasmid...plasmid page also helps researchers find other publications that have used the same plasmid through the ...section. Having IDs linked to these metadata and publications help to enable reproducibility of experiments...and N2T.net. To help authors comply with journal policies, Addgene now provides an RRID and a resolvable... in the material and methods section of your publication. You should carefully read and follow the journal...

...2020.  “CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future.” Front. Oncol... for full details.    CRISPR advances in monoallelic diseases In 2013, a team led by Jinsong Li from...the technology in different labs for different applications.” Li’s team noted that CRISPR/Cas9 has been ...with size limitations. In addition, non-viral applications are also possible through the use of nanoparticles....  “CRISPR Genome Editing Technology and its Application in Genetic Diseases: A Review.” Current pharmaceutical...

...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... science? Or perhaps you have presented in some public lectures? These transferable skills could be meaningful...make their job postings and salary expectations public, so start by looking at these university postings...

... our knowledge. Disease Applications Q13: Does AAV have potential applications in diseases which may be... to use split-inteins to mediate trans-protein splicing. Note, however, that this will require co-delivery...reliable and reproducible. However, we do not use amplicons in ITR as we have found this to strongly bias ...1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011 - 1x1012 vg/...CRISPR tools is an exciting area for future AAV application to epigenetic editing. This area is discussed...

...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...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...

...protein for your application. Selecting the best fluorescent protein for a specific application can seem like...under conditions that best mimic the intended application. The three key properties for live cell imaging...experimental set up. The practical brightness takes all application specific parameters into account, including the... picture of what can be expected in a 'real' application than the theoretical brightness. Consequently...brightness in a system closely mimicking the future application. In figure 2 we provide an example of the comparison...

...transcription, or DNA repair, or replication, or any number of CRISPR applications and thought “OK, but how does...with Addgene, but you’ll find many great tools by clicking on the links below and visiting their associated...containing lists of plasmids from the indicated publications. You can request all of these plasmids directly..., Hoefler R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen ..., Hoefler R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Rapid and...

...tool that can be used for a variety of research applications. Within the immune system, antibodies are produced...antibodies to bind proteins is useful for research applications as well because they allow scientists to target...150 kDa, a nanobody is 12-15 kDa.   Research applications for antibodies You can use antibodies for experiments...Let’s dive in and take a look at some of these applications. Western blot Western blots are used to detect...immunocytochemistry). This could be cells or tissue slices mounted to a slide, or whole organs. Because the... by binding to cell surface receptors. These applications are typically used for investigating signaling...are out there, and how many different research applications they can be used for. Addgene’s Antibody Plasmid...

...ampicillin resistance cassette and an origin of replication, which are not schematized. Sizes of fragments...2024, used under the Creative Commons CC BY-NC-ND license.   Once you have the components you want, simply...validated and were chosen because of their broad applicability in different model systems. What’s the error...five months. When they tested those clones (in duplicate), 93% had the correct banding pattern in the restriction...correct. Because they created the assemblies in duplicate, they were able to confirm that at least one plasmid...design. Note that you can browse all components by clicking the blue text in the upper right-hand corner.  ...

...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no...technology park, it enjoys preferential governmental policies for a high-tech development zone so many science...inheritance that might be less error prone than DNA replication. He too mentioned how “surprising” small RNAs...created a circRNA website so scientists can view public circRNA datasets and download the custom python...Steitz quipped that RNA in the ribosome is in a complicated conformation “not like DNA which has a boring...

...are, you’ll even have DNA left over for other applications, like PCR, cloning, transfection, or long-term...wash step or use a different strain. For most applications, Addgene uses and recommends DH5𝝰! This strain...your E. coli to ensure it is suitable for your application. We have previously reviewed many common lab ...partially specified by a plasmid’s origin of replication, but can also be affected by the plasmid size...the material used for inoculation. To further complicate matters, plasmid copy numbers are often ambiguous...volume limits, the copy number of some origins of replication (e.g. p15A, ColE1) can be increased by adding...