Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 839 results for: LEA

Showing: 161 - 200 of 839 results

  1. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    Published
    Dec. 3, 2020, 2:15 p.m.
    ...its endonuclease activity to generates a ssDNA substrate which is then nicked by its endonuclease domain...engineered Learn about the parts of the AAV transfer plasmid  Resources on Addgene.org Learn about AAV...of Rep78 and Rep68 contains a DNA-binding and endonuclease domains, all four Rep proteins share a common...virion assembly, the role of MAAP has not been clearly defined yet. Now you’ve got a basic understanding...AAV RepCap plasmid! Check out this blog post to learn more about the AAV transfer plasmid! References...

  2. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    Published
    June 4, 2020, 1:22 p.m.
    ...depurination, the hydrolysis of the glycosidic bond leads to the release of a purine (adenine or guanine) from the...DNA are repaired by base excision repair, but can lead to mutations in single stranded DNA undergoing replication... from E. coli is the presence of the protein endonuclease I encoded by the gene endA. Nowadays, the most...gene has been removed/mutated to neutralize the nuclease activity. For strains that still carry this gene...introduced in the purification protocol to remove endonucleases activity. For these reasons choosing the correct...most downstream processing. On the other hand, nuclease free water is a great option because it provides...

  3. Addgene's a Nonprofit? Nonprofit Awareness Day 2017

    Type
    Blog Post
    Published
    June 5, 2017, 3:19 p.m.
    ...professional Addgene is a thought leader in scientific sharing, leading by example to increase sharing in...Resources on Addgene.org Learn More About Our Mission Meet the Addgene Leadership Team Get the Latest News...arts organization. Many people are surprised to learn that Addgene is officially filed, recognized and...is a nonprofit? The term "nonprofit" is a bit misleading. Nonprofits can generate revenue (as Addgene ... Xuan Additional Resources on the Addgene Blog Learn about Our Collaboration with Fellow Nonprofit Seeding...

  4. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

    Type
    Blog Post
    Published
    March 15, 2018, 12:59 p.m.
    ...By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create...mutation inactivates the RuvC domain, so this nickase cleaves only the target strand. Conversely, the H840A mutation... in the HNH domain creates a non-target strand-cleaving nickase. Instead of cutting both strands bluntly...distances (40 - 130 nt). From these results, it’s clear that editing is much higher in the PAM-out configuration... At the AAVS1 locus, the two nearest gRNAs had cleavage sites 12 nt and 13 nt away from the target site...wide margin, exhibiting >20% repair efficiency. Clearly, a nickase strategy can expand the targetable region...post. Additional Resources on the Addgene Blog Learn about Homology Directed Repair 3 Tips to Improve...

  5. Oh, The Places You Can Go: Careers in Science Communication - Product Development

    Type
    Blog Post
    Published
    Oct. 18, 2017, 1 p.m.
    ...Careers in Science Communication blog series, you’ll learn about Caitlin Runne-Janczy, a Product Development...While her love of educational science writing is clear, Caitlin’s career plans weren’t concrete when she...what I wanted to do, but knowing that I wanted to leave the bench.” Her path to figuring out her career ...writer.” At the end of her postdoc, Caitlin had a clearer idea of her career goals: “My plan was to go into...for part two of Caitlin’s interview where you can learn what it’s like to work for a science education company...series!  Additional Resources on the Addgene Blog Learn more about alternative science careers here Be like...while in grad school Resources on Addgene.org Learn more about careers at Addgene ...

  6. Oh, The Places You Can Go: Careers in Science Communication - Science Writing

    Type
    Blog Post
    Published
    Oct. 3, 2017, 12:11 p.m.
    ...non-academic career Learn more about alternative science careers here Resources on Addgene.org Learn more about...series was written for selfish reasons: I wanted to learn about careers in science communication. When I started...away-from-the-bench career option. So if you’re interested in learning more about science communication careers, you’...was a science writer, reached out to me. She was leaving a writing position and looking for someone who ...s paid off. Work is never boring. “I just keep learning on the job. It keeps it interesting.” And while...topic, but I haven’t met any.” A few other tips: “Learn HTML, write a lot to make sure you don’t hate it...

  7. Addgene Begins Distribution of Recombinant Antibodies

    Type
    Blog Post
    Published
    March 28, 2022, 4:30 p.m.
    ...s antibodies are plasmid-based and recombinant. Learn more about recombinant antibodies here. Finally,...Finally, Addgene is a nonprofit that connects colleagues and is invested in improving research infrastructure... to maximize the reach and impact of your work. Please email [email protected] if you would like to ...plasmids via Addgene! Email [email protected] to learn more or get started.  If you have suggestions about...ideas for new materials to add to the collection, please let us know by filling out this suggestion form... will find useful antibodies for your research. Please consider sharing your experiences with antibodies...Resources We would also like to acknowledge our colleagues at the Developmental Studies Hybridoma Bank, ...

  8. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ...Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts in your experiments. Read this ...Professor at Albert Einstein College of Medicine, to learn how problematic oligomerization can be and how to...that occur during development. Read this post to learn more about recombinase-based state machines and ... Green (both formerly at the Wyss Institute) to learn how these amazing tools may be used to rapidly diagnose...you can use to study basic virus biology while leaving virulent components behind. Minigenomes are stripped...and transcription as well as reporter genes. To learn how you can use minigenomes in your research, check...

  9. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ...nature of non-homologous end joining (NHEJ) often leads to the generation of indels and thus frameshifts...activation) and CRISPRi (inhibition) technologies – a nuclease-dead Cas9 (dCas9) is directed near the promoter...landscape of mismatches that can nevertheless still lead to activity has been established, and can be used...that the consequences of off-target activity, at least for the experimental conditions used, led to no ...the generation of truly isogenic cell lines than nuclease-mediated off-target effects.” Further, large-scale...al., 2019). Again, the experimental strategy is clear: for any gene of interest, one should require that... Liu DR (2020) Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors....

  10. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...the Addgene Blog Learn more about FRET Tips for Using FRET in Your Experiments Learn more about Fluorescent...brightest since it had an emission near 600 nm – a clear demonstration of why a BRET reporter can improve...Many intracellular sensors of events like calcium release have been made using fluorescent resonance energy... molecules. FRET sensors face challenges of photobleaching, autofluorescence, and, in the case of exciting...wavelengths, excitation of the optogenetic regulator leads to spurious activation of the FRET sensor. Researchers...et al. (7) wished to trigger intracellular Ca++ release with an optogenetic receptor, melanopsin, requiring...#83926). Live-Cell Voltage Sensor  Ingaki and colleagues (8) were interested in using photoactivatable...

  11. Finding Your Perfect Job After University

    Type
    Blog Post
    Published
    Jan. 12, 2016, 3:30 p.m.
    ... about roles in this field, most jobs needed at least 3 months experience working in a developing tropical...positive experience. I enjoyed helping people and learning to explain concepts in ways that people from another...traditional medicine couldn’t fight effectively. I had to learn how to cope with a different climate and culture...work became monotonous very quickly; as soon as I learned how to do the different tests and became efficient...where one drug does not work for all patients, leading to very high mortality. I really loved the fact...cannot be as adaptive. I was also surprised to learn that I enjoy the customer service side of my job...vital because otherwise I’d lose touch with my colleagues and have misunderstandings.  Working long distance...

  12. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 14, 2019, 1:54 p.m.
    ...describe a process that begins with cell lysis to release AAV particles with subsequent purification steps...also possible for AAV vectors to be spontaneously released from the producer cells into the supernatant during... are exosome AAVs? In 2012, Casey Maguire and colleagues from Massachusetts General Hospital and Harvard...took a closer look at the AAV vectors that are released from cells during production (Maguire et al., ...the exo-AAV isolation protocol by Maguire and colleagues, other researchers have tried to modify it to...possibly improve transduction properties. György and colleagues found that the association of AAV particles with...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

  13. Science Career Options

    Type
    Blog Post
    Published
    Feb. 23, 2017, 1:30 p.m.
    ...often a hard question to answer if you plan on leaving academia, but don’t quite know what you do want...goal, and when you realise that your goal might not lead to a career you actually want, you can feel lost...Journal Editor Science Marketer Software Developer Learn Science Communication Skill from a Student Group...in the media and in films, which can often be misleading. Key aspects to determine are working hours (... the job require public speaking? Will you be leading a group or working as part of team? Will you be...application. Additional Resources on the Addgene Blog Learn How to Experiment with New Careers while in Grad...Koeris' Journey from Grad Student to Entrepreneur Learn to Network and Make the Contacts for Your Dream ...

  14. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    Published
    Feb. 9, 2021, 2:15 p.m.
    ...miscommunication, a lack of leadership, and inappropriate tools or writing approaches can lead to frustration, delay...Weill Cornell Medicine. Discussing data with colleagues can be more difficult during the pandemic era...can contribute and sketch on the board without leaving Zoom. The annotate function can also make the screen-sharing...cultural backgrounds.” This work outlines the need for clear communication, fair credit through authorship, and...ways of working, reconnect as human beings and colleagues, eventually encouraging purposeful and creative...Medicine in NYC and author of the book “The Salmon Leap for PhDs: Swimming upstream a career from academia...

  15. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    Published
    Dec. 10, 2019, 1:58 p.m.
    ...bound lipoproteins Article contributed by Leah Schwiesow A leading cause of death worldwide, cardiovascular...ever like to write about a recent plasmid deposit please sign up here.        A retroviral mouse CRISPR...genetically. To solve this issue, Sarah Teichmann and colleagues created a retroviral CRISPR knockout library ...lipoproteins through the circulatory systems, but can also lead to the formation of atherosclerotic plaques. ApoB...through triglyceride/cholesterol does not provide a clear view of either of these metrics (Sniderman et al...-LP size and concentration, Steven Farber and colleagues at the Carnegie Institute for Science have developed...

  16. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    Published
    Nov. 7, 2019, 2:18 p.m.
    ...of a paper is the one that you need to spend the least time on—just put down a basic outline and refer ...figure them out. But, when they come to light, it’s clear that what the researchers thought was a problem ...role, and we take our responsibility to push for clear communication of science very seriously. That’s ...and transparency of methods sections, including clear information about reagents, and why other publishers...want to be? The part that is still missing is a clear alignment of incentives. Right now, researchers ...evidence of sharing for grant renewal—but there is clearly room to do more. If we could really instill a culture...Her current responsibilities encompass overall leadership of the Cell Press editorial department, a team...

  17. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ... we've heard over and over from our users: "Please, please improve your plasmid maps!" After thoughtful... here to see the new map in action). Read on to learn more about the improvements we’ve implemented and...extraneous ORFs that cluttered our old plasmid maps. Clearly Annotated Features, Enzymes, and Primers - The ...easier for you to share plasmid information with colleagues, or explain plasmid features to a lab member ...Once you’ve identified a plasmid that you want to learn more about, updates to our sequence analyzer will...hover over any feature, enzyme, primer, or ORF to learn more information about it. For example, if you hover...

  18. A Deep Dive into BioBuilder

    Type
    Blog Post
    Published
    Oct. 11, 2017, 1:44 p.m.
    ...through October 19. The BioBuilder Learning Lab The BioBuilder Learning Lab is a fully equipped, state-of-the-art...and teachers in the U.S. and beyond. “I want to learn more about Synthetic Biology. Can you tell me where...capitalize on the excitement found through experiential learning. “Students may forget the technical details they... BioBuilder experience With its intentionally malleable curriculum, BioBuilder takes cutting-edge research...a project-based curriculum, BioBuilder expands learning outcomes by emphasizing the importance of critical...production of fire-resistant housing insulation, and cleaning up salt runoff from winter icing.BioBuilderClub...

  19. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    Published
    July 10, 2014, 7:17 p.m.
    ... Sample6 has changed focus and names, swapped leadership, and grown to 25 members.  What is Sample6?...competing in business plan competitions. After much learning, the focus of the application was changed, but...best, wins. He explained that entrepreneurs must clearly communicate their idea at various levels of complexity...industry vary drastically, he reassures that one can learn it by reading a broad variety of materials and constantly...million. The biggest challenge during this time was learning to communicate effectively to non-scientific audiences...are essential to the biotech startup puzzle: Nucleation of the team. The team is key, and the initial...raise money and/or validate an idea or product? Please let us know in the comments section! And read on...

  20. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase activities.    ...site for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from its terminal ...terminal sequences at the excision site after cleavage by the transposase. It has a cargo capacity >100 kB, although... TTAA and unlike other transposons, it does not leave behind a DNA footprint sequence after excision. ...   Applications of transposons Now that you’ve learned about some of the popular transposon systems, let...DNA into the pronuclei of fertilized eggs, which leads to random incorporation of that sequence into the...ijms22105084 Additional resources on the Addgene blog Learn about the INTEGRATE system for bacterial genome ...

  21. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...the RNases on your hands can degrade RNA. Keep a clean work area, which may include spraying your bench...Alternatively, glassware can be baked at 180°C for at least 4 hours.   If your final RNA sample(s) are resuspended...cell lysis.  In the late 1970’s, Chirgwin and colleagues showed that a strong protein denaturant, guanidinium...mindful of your extractions- if you need really clean RNA, it’s important to make sure that when extracting...effect of ammonium sulfate on the activity of ribonuclease A. Biochimica et Biophysica Acta (BBA) - Enzymology...active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299. https://doi.org/... Leah Schwiesow...

  22. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...macroautophagy of the cytosol leads to accumulation of Pho8Δ60 in the vacuole, where it is cleaved to generate an active... molecular weight shift of Pho8 from its uncleaved to cleaved form is the final read out of this assay...cytosol, to the higher pH of the vacuole. Read on to learn more about prior methods for studying autophagy ...the vacuole via the secretory pathway, and then cleaved to generate an active form of the protein. To monitor...lysosome, since both Atg8p and LC3 are degraded or released once an autophagosome fuses with the vacuole or...

  23. Developing Lab Management Software for Biology

    Type
    Blog Post
    Published
    Aug. 25, 2016, 2:30 p.m.
    ...requisite for a functioning lab, but it is for a scaleable lab. When you need to track the location, quality...post we’ll highlight some of the lessons we’ve learned over the years with the hope that our insights ...containers and contents Right from the get-go be as clear as you can be (there will always be confusion about..., tightly coupling them to their contents might leave you sad and confused when a tricky procedure failed...worm that requires a change of tube, there’s no clean way to separate the contents from the container ...etc.There are also going to be situations where you learn biology terms (plasmid, 5-prime, CRISPR, growth ...krisshamloo.com     Additional Resources on the Addgene Blog Learn about How We Use Barcodes in the Lab Get Tips on...

  24. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    Published
    July 21, 2016, 2:30 p.m.
    ...the Addgene Blog Learn How Scientists Are Trying to Shape the Future of Research Learn How Notions of Scientific...studies with concrete examples of how gender analysis leads to discovery and innovation. One such case study... of their cells when doing experiments—which can lead to failed research. For example, an international...develop represent the whole human population. To learn a little more about where differences in effects...There is much work to be done! Researchers need to learn sophisticated methods of sex and gender analysis...Blogger Londa Schiebinger. Londa Schiebinger is a leading international expert on gender in science and technology...

  25. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    Published
    Jan. 29, 2015, 1:25 p.m.
    ...Ask openly for clear information on costs. Can you suggest some coaches? Yes I can. Please note that Addgene...to change jobs? Grow your professional presence? Learn a key new job skill? Finish your thesis? Knowing...that will help you. A good coach usually will be a clear communicator and make direct suggestions for improvement...She is also a very popular speaker and workshop leader. Lauren Celano of Propel Careers is not a professional...engagements and has many areas of specialty from leadership development to mentoring for women and under-...Guide Your PhD with An Individual Development Plan Learn a Little about The Academic Labor Environment from...

  26. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...mEosFP exposure to UV light causes an irreversible cleavage near the chromophore, resulting in emission of...to blue light also inactivates the chromophore, leading to a non-fluorescent state. Dronpa can be reversibly...quickly inactivated and less photons are emitted leading to dimmer fluorescence. In fact, many of the early...are being emitted, making it easier to capture a clear image above background. The majority of PA-FPs come...disrupting cellular function and localization, or leading to protein aggregation. Another important parameter...light used to activate the first PA-FP does not photobleach the second, and vice versa. Finally, as every...contrast, PA-FPs offer a reduced potential for photobleaching and phototoxicity that standard FPs might induce...

  27. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    Published
    July 11, 2019, 1:26 p.m.
    ...implanted into a female animal for gestation. Nucleases that induce double-stranded DNA breaks, such as...cells, which are then injected into a blastocyst. Nucleases can also be used to engineer knockout mice. ZFN...non-homologous repair at the site of DNA damage. These nucleases are microinjected into a zygote, which is then...introduce point mutations. While using targeted nucleases to generate an engineered mouse model has advantages...targeting vector alone, some challenges still exist. Nucleases may cause off-target effects on regions of the... random process, zygotes edited using the same nuclease may not result in genetically identical mice. ...about Cre-lox Read our Genome Engineering Guide Learn more in our CRIPSR guide ...

  28. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ... 2019). Inhibiting DNA cleavage Example: AcrIIC1 binds to the HNH endonuclease domain of Cas9 and prevents...prevents target DNA cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR proteins (Acr's) can...loading, inhibiting DNA binding, or inhibiting DNA cleavage.   Five ways to anti-CRISPR proteins in your...timing experiments, the Doudna lab found that at least 50% of on-target Cas9 edits happens within the first...effects and cytotoxicity that result from excessive nuclease activity in tissues (Li et al., 2018). 4. Selection...Blog First time CRISPR user? Check out this post. Learn how to Choose the Best Cas9 Variant for Your Next...

  29. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...anymore. Say hello to PAM-free and PAM-flexible nucleases! In this blog, we will review Cas enzymes, which... uses a PAM with no sequence constraints, or at least has looser or different requirements. Luckily, there...Nonetheless, in an AT-rich region, these Cas9s may still leave you choosing between efficiency and location.   ...display higher off-target effects than SpCas9, a pleasant surprise since the PAM flexibility could in theory...activity of SpRY was validated in double stranded cleavage and base editing applications, with high editing... NG Cas9 NG - SpG Cas9  NG  - SpRY nuclease  C to T Base Editor SpRY A to G Base Editor SpRY...  Nishimasu, H., et al.,Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science 361,1259...

  30. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...that delivering RNA-guided nucleases or TALENs together with AdV donor DNA leads to a vast majority of AdV-modified...chromosomal region. The CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various... and demonstrated by Manuel Gonçalves and his colleagues at Leiden University Medical Center are now available...DNA for accurate genome editing with engineered nucleases. Nat Methods 11:1051–1057 . https://doi.org/10.1038...Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse...delivery of CRISPR/Cas9. Journal of Controlled Release 298:128–141 . https://doi.org/10.1016/j.jconrel...

  31. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...DNA could lead to transgene expression in recombinase-negative animals, producing misleading results.  ...the plasmid used to create the viral prep. In a clean viral prep, like AAV-44362 shown below, more than...from these known impurities, as was the case for clean sample AAV-44362. In addition to these expected ...number of hits to the same sequence (gene). In a clean sample, there are typically fewer than 10 hits to...receive the best AAV vectors for your research. Please use the comments section below to let us know if...Topic Page Subscribe to Viral Vectors Blog Posts Learn How We Use NGS for Plasmid Quality Control Resources...

  32. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...This DNA degradation portion can be bypassed by leaving out Cas3 — a tactic some CRISPR transposase systems...chop DNA. Cas3, however, has two jobs, one as a nuclease and one as a helicase. Following recruitment by...initial steps, Cas3 can perform its helicase and nuclease functions simultaneously, by continuously unraveling...unraveling dsDNA, pulling through the ssDNA, then cleaving the ssDNA strands. In this way, Cas3 eats away... into think it’s more powerful than it is. The nuclease domain of Cas3 is considered weak, especially ...However, by continually passing the ssDNA past the nuclease domain, Cas3 is able to repeatedly cut the DNA...thymine-rich regions on the non-target strand. Occasional cleaving of the target strand is not as well understood...

  33. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...including mitosis, mitochondrial fission/fusion, nuclear import, and neuronal trafficking. Although FPs ...Overexpression of fluorescently tagged proteins can lead to improper protein localization, protein aggregation...power of this system lies in the ability of the endonuclease Cas9 to create a DNA double stranded break (...the tag, selectable marker, and connector to be released (Fig. 1, Step 2-3). Since donor plasmids of the...directed repair after a CRISPR/Cas9 induced break. Learn more about these constructs and the cell lines they...Cas systems used for endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein...PMC4889673. Additional Resources on the Addgene Blog Learn about Fluorescent Protein Timers Check out Photoactivatable...

  34. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ... PITCh plasmid to release the cassette, and an insertion-locus-specific gRNA cleaves genomic DNA. These...it’s become clear that the power of this editing platform lies in the diversity of nucleases and their .... For more mechanistic details on HDR and NHEJ, please see the linked blog posts. Compared to its counterparts...compare to CRISPR sticky-end insertion using the nuclease Cpf1. Since Cpf1 cuts in a staggered pattern, ...Experiment Additional Resources on Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find ...

  35. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ... Developed by David Liu and colleagues, FokI-dCas9 expresses Fok1 nuclease domain fused to catalytically... to this system by engineering cells that lack release factor one (RF-1; Bacterial strain EcAR7) and minimizing...control for your experiments. For more information, please see Addgene’s information page for the Rinehart...neuronal signaling. For more information on DREADDs, please see the Roth lab's DREADD users blog. New ... expression plasmid that was used by Chen and colleagues for genome editing inXenopus tropicalis. (Guo...information on using the Platinum Gate TALEN Kit, please see the Yamamoto lab’s protocol for TALEN construction...mGRASP A Nature 2012 article by Jinny Kim and colleagues describes their efforts to map the location and...

  36. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ...editors. They adjust the side groups of DNA bases and leave the core ring structure intact, allowing them to...guide RNA to base pair with the DNA target strand, leaving the non-target strand unpaired and accessible on...it the “edited strand.” Uracil excision repair leads to the first base transversion editor The story ...improved the CGBE: DNA polymerase D2 (POLD2), exonuclease 1 (EXO1), and RNA binding motif protein X-linked...included in the CGBE. They also developed a machine learning model capable of predicting editing outcomes of...that were already accessible. It's not entirely clear why this is, but it might point to a more dynamic... screens, target-library analysis, and machine learning. Nature Biotechnology, 39(11), 1414–1425. https...

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

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ...to see the dirty laundry exposed here, but it’s unclear how much they will actually care. The section of...technology. Though these case studies are “old,” they lead Zimmer to one of his most critical observations:...detection had over 1,000 authors. The chapter on deep learning addresses the dilemma of creating a technology...discussed The State of Science with the five Addgene colleagues who read it with me, the consistent response ...non-scientist alike was that the book left them wanting to learn more. There are individual paragraphs that may have...task to set oneself. As a scientist who enjoys learning a little bit about many different things (part...about a long time. For the scientists who like to learn everything about one specific topic, The State of...

  38. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...several washes, this tag can be cleaved by the bdNEDP1 protease, thus releasing purified nanobodies from the...common protein purification methods. Read on to learn more about nanobodies and how their structure and...primary antibodies raised in different species, or at least from different IgG subclasses, so that secondary... and the target antigen to about 2 nm, which can lead to higher resolution for super-resolution microscopy...rabbit nanobodies. If you’re interested, you can learn more about how nanobodies are identified in this...University Use Nanobodies to Control Protein Activity Learn more about protein tags Read about E. coli strains...

  39. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ... clinical gene therapy applications. Read on to learn more about the current (and future) applications... co-transfection of these plasmids, 293T cells released transgene-containing lentiviral particles into...and they showed that expression persisted for at least thirty days. Although adeno-associated viral vectors...activation from the 5’ LTR) and Rev (facilitates nuclear export of transcripts). A different viral envelope...safer, this system may be more cumbersome to use and lead to lower viral titers due to higher plasmid number...Addgene Blog Your Lentiviral Plasmid FAQs Answered Learn How Lentiviral CRISPR Libraries Enable Genome-Scale... on Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors for...

  40. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...journals. In addition, it is my pleasure to interact with my colleagues in meetings of scientific societies..."Biology is Method", i.e. new technologies often lead to big discoveries in biology. Therefore, my lab...translocation trap (ITT) system to analyze signal-induced nuclear translocation of proteins in 2004. In addition,...much enjoyed interactions with many excellent colleagues at NYU and visiting scientists, such as seminar..., I've been very lucky to work with excellent colleagues in my lab. On the other hand, I feel that researchers... conventional research. So, I hope my younger colleagues may be able to develop new technologies, hopefully...
Showing: 161 - 200 of 839 results