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  1. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...Nanobodies from camelids have gotten a lot of press for being a promising approach for treatment or prophylaxis...The Seeger lab found a way around this by using plasmids to generate sybodies, synthetic nanobodies against...nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target epitopes that...fewer hydrophilic surfaces, require detergents or lipids to remain folded, and are often less abundant than...trinucleotides (corresponding to different amino acids) in the regions that bind to the antigen. The Seeger...form of mRNA for academic research. The template plasmids for generating your own libraries are also available... restriction sites (Geertsma, 2013). Once the phagemids are constructed, they are used to produce a phage...

  2. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...three plasmids — a transfer plasmid containing your gene of interest plus two packaging plasmids (three...three packaging plasmids for 3rd generation lentivirus production). The packaging plasmids are widely available...bacmid. Like AAV and lentiviral packaging plasmids, bacmids are readily available, so the amount of cloning...about cloning your transfer plasmid. The transfer plasmids have different packaging capacities depending ...plasmid. Production of baculovirus requires only two plasmids — your transfer plasmid and the baculovirus construct...insect and mammalian systems. Baculovirus transfer plasmids have a much larger packaging capacity — up to ...can produce AAV and lentivirus if needed # of plasmids Three or four Two Packaging limit ~4.7 kb (AAV...

  3. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...especially for the study of pain. While many new capsids (i.e. the part of the virus that determines tropism...Caltech stepped up to the challenge. Find the PHP Plasmids Here! Their Solution: Use the CREATE method to... to engineer two new IV injectable AAV capsids, AAV.PHP.eB and AAV.PHP.S, that target the CNS or the PNS...  Read on to learn more about these two new AAV capsids and how they can be useful tools for studying the..., a pool of mutant AAV capsid (cap) expressing plasmids is generated. Diversity is introduced into the...Delivering the CREATE AAV library via IV screens for capsids that are able to cross this barrier - only those... do these AAVs transduce the CNS and PNS? Both plasmids infect the PNS or the CNS much better than the...

  4. The MTA at Addgene

    Type
    Blog Post
    ...organizations choose to share their plasmids under an OpenMTA, which allows plasmids to be used for both research...If you’ve deposited or requested plasmids from Addgene, then you’ve been involved in the material transfer...Even though this process is essential to sharing plasmids around the globe, it can be a bit of a mystery...use agreements. The UBMTA, which is the MTA most plasmids at Addgene are shared under, is well-understood...we are not a party (signatory) to the MTAs for plasmids in our repository. Instead, we facilitate the ...institutes. But they’re not! The vast majority of our plasmids are distributed under the UBMTA, which is widely...share your biological materials under! While most plasmids at Addgene are shared under the UBMTA, researchers...

  5. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ...Chromatin-related genes with plasmids available from Addgene The links below pull up lists of plasmids containing the...containing lists of plasmids from the indicated publications. You can request all of these plasmids directly from...study chromatin and want to start sharing your plasmids through Addgene, don’t hesitate to start the process...these pages to learn when Addgene receives new plasmids containing these genes. As with the PIs above,... In-exhaustive Chromatin-related genes with plasmids on Addgene.org AGO1 BRCA1 CREBBP CUL1 H2AFX...) ARID1A   Chromatin research articles with plasmids available from Addgene Many articles reporting...chromatin-related research describe the production of plasmids that might be useful for your work. The links ...

  6. Addgene: The First Twenty Years

    Type
    Blog Post
    ...frustrated by the difficulty of accessing published plasmids from other scientists. She, Kenneth Fan, and Benjie...nonprofit that would facilitate the sharing of plasmids between scientists. The service would allow people... our first major repository milestone — 10,000 plasmids deposited and available — and nearly seven before...possible for scientists to both deposit and request plasmids, was a major focus of our first decade. We did...work In March 2015, Addgene reached the 500,000 plasmids distributed milestone! We were thrilled to see...also made us think of all the depositors whose plasmids we were entrusted with. So in June 2016, we launched...community. More than 100 labs worldwide use our plasmids. I hope they contribute to great discoveries!”...

  7. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...AAV1 and AAV2, with only six amino acids distinguishing its capsids from those of AAV1 (Issa et al., 2023... true! In a previous post, we covered systemic capsids, which allow AAVs to broadly spread throughout ...BioRender.com. AAV vectors have both distinctive capsids and genomic structures. The viral capsid is the...your choice of promoter/enhancer. Table 1: AAV capsids and receptors. Capsid Primary receptor Coreceptor...Coreceptor AAV1 α2,3-/α2,6 N-linked sialic acids1 AAV receptor (AAVR)2   AAV2 Heparan sulfate ... α and β3 AAV6 α2,3-/α2,6 N-linked sialic acids1 Heparan sulfate proteoglycan (HSPG)3 AAV receptor...nomenclature (AAV1, AAV2, etc.) to distinguish the capsids from one another. Common serotypes We’ll cover...

  8. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...elegans Browse our Plasmids 101 blog posts Resources on Addgene.org Visit our worm plasmids and resources...discussed at the meeting, in addition to useful plasmids and toolkits available at Addgene.  C. elegans... on new resources and tools for the community. Plasmids for the Auxin Inducible Degradation System (AID...and temporally knock-down proteins of interest.  Plasmids for cGAL and split cGAL systems The Sternberg ...single tube. You can also find additional SapTrap plasmids used for introducing point mutations that was ... additional resources about C. elegans and the plasmids we have at Addgene check out our worm expression...continuously curated as we continue to receive plasmids designed for the worm so check back frequently...

  9. Pooled Library Amplifications

    Type
    Blog Post
    ... overtake full-size plasmids (or for smaller plasmids to overtake larger plasmids, if the library varies...How many unique plasmids does your library contain?), the variance of size of plasmids in your library...30 to 200 times as many colonies as individual plasmids in your library. As an example, if you want 100x... your library, and your library contains 5,000 plasmids, your goal for your transformation would be to...libraries often contain thousands, if not millions, of plasmids, they are generally electroporated into electrocompetent...based on the size of and weight of your library plasmids. As long as you achieve sufficient coverage from... Addgene.  Library Type Name Number of Plasmids Variable Region Size DNA per Amplification...

  10. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    ...feature some recently deposited plasmids every quarter. The “hot plasmids” from last March were quite popular...past year: CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These...s the top 10 blog posts from the year. 10. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent...of tools we share on behalf of scientists. 9. Plasmids 101: Positive and Negative Selection for Plasmid... about mNeonGreen, mTFP1, and mWasabi and find plasmids containing these fluorescent proteins at Addgene..., and all possible base-to-base changes.   1. Plasmids 101: Transformation, Transduction, Bacterial Conjugation...Alyssa Cecchetelli The introduction of nucleic acids into bacteria or eukaryotic cells is a common technique...

  11. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...is mediated by ADAR2DD, not PspCas13b. Find the plasmids for the RESCUE RNA editors here! Testing and improving...for viral delivery.  Find the RESCUE RNA editing plasmids here! Future directions Building on previous RNA...Science (2017):pii: eaaq0180. PMID: 29070703. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...550(7675) (2017):280-284. PMID: 28976959. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...27256883. PubMed Central PMCID: PMC5127784. Find plasmids from this paper at Addgene. East-Seletsky, Alexandra...) (2016):270-273. PubMed PMID: 27669025. Find plasmids from this paper at Addgene. Gootenberg, Jonathan...28408723. PubMed Central PMCID: PMC5526198. Find plasmids from this paper at Addgene. East-Seletsky, Alexandra...

  12. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    ... 1998). Interested in plasmids that contain the groucho gene? Check out plasmids deposited by the Shivdasani...Drosophila Read our Plasmids 101 blog series Resources on Addgene.org Find plasmids for your research ...the stories behind the names. We also have many plasmids in our repository that you can use to study these...right and left hemisphere. Addgene has several plasmids that contain SHH, you can find them all here! ... Beauty (Ivics et al., 1997). We have a lot of plasmids that contain components of the SB transposon system...actin fibers that resemble spaghetti. We have two plasmids in our inventory from the lab of Eric Wieschaus...different animals. Check out the Addgene website for plasmids that contain tinman or the mammalian homolog Nkx2.5...

  13. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...into adeno-associated virus (AAV).  At 984 amino acids in length, Cas9 from Campylobacter jejuni (CjCas9...284 . https://doi.org/10.1038/nature24049 Find plasmids from this paper at Addgene. Amrani N, Gao XD,... https://doi.org/10.1126/science.aaq0180 Find plasmids from this paper at Addgene. Kim D, Kim J, Hur...863–868 . https://doi.org/10.1038/nbt.3609 Find plasmids from this publication at Addgene. Kim E, Koo ...8: . https://doi.org/10.1038/ncomms14500 Find plasmids from this publication at Addgene. Kleinstiver...485 . https://doi.org/10.1038/nature14592 Find plasmids from this publication at Addgene. Kleinstiver...869–874 . https://doi.org/10.1038/nbt.3620 Find plasmids from this publication at Addgene. Hu JH, Miller...

  14. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... new plasmid. One area with lots of bright new plasmids is our Fluorescent Biosensors Collection — scientists...SF-iGluSnFR. Image from Aggarwal 2022.   Find iGluSnFR3 plasmids here! Find viral prep here: pAAV.hSyn.FLEX.iGluSnFR3.... 2022, under CC-BY-NC license.   Find JEDI-2P plasmids here! Find viral preps here: pAAV-EF1a-DIO-JEDI...et al. 2022, under CC-BY license.   Find pYtags plasmids here!   Farahani PE, et al. pYtags enable spatiotemporal... suitable for AAV packaging.    Find iGECInano plasmids here!   Matlashov ME, et al. Design and Initial...CC-BY-NC-ND license.   Find KRaION and Kbp sensor plasmids here!   Torres Cabán CC, et al. Tuning the Sensitivity...et al. 2022, under CC-BY license.   Find LiLac plasmids here!   Koveal D, et al. A high-throughput multiparameter...

  15. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    ...blogger Joe Mellor, Founder and CEO of seqWell Inc. Plasmids and PCR products are the bread and butter of molecular...Addgene’s large and growing collection of curated plasmids from all over the world. The rest of this piece...partnership with Addgene to sequence large numbers of plasmids. plexWell™ technology overview For any NGS experiment...individual input DNA samples (e.g. a collection of 96 plasmids on a plate) will typically have a wide range of...to, for example, assemble hundreds of different plasmids from a single Illumina MiSeq® run. As shown in...plexWell™ + Addgene = high quality NGS-assembled plasmids Because of the enhanced normalization of read ...large-scale de novo assemblies of short constructs like plasmids. The plexWell™ workflow also offers a highly efficient...

  16. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    ...continue having access to plasmids during the COVID-19 pandemic. Over 7,000 plasmids for COVID-19 research...work to share plasmids to all scientists Resources on Addgene.org Find COVID-19 plasmids at Addgene Browse...Biological repositories that distribute mouse strains, plasmids, cell lines, and much more have been critical ...science is truly a global endeavor. Case study 2: Plasmids Another example of reagents that have broad applications...(Ou et al., 2020). The authors requested these plasmids from Addgene. This publication then resulted in...requested from Addgene. Figure 2: Total COVID-19 plasmids requested from Addgene from March 23, 2020 to ...to July 27, 2020. Browse Addgene's COVID-19 plasmids and resources! Case study 3: Mouse strains Reagents...

  17. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...light-responsive system is spread across three plasmids, with the response regulator encoded in the genome...et al. condensed the system to two streamlined plasmids. The utility of these TCSs was limited by a few...These bulky systems were spread over a number of plasmids, with one RR (ompR) encoded chromosomally. Leakiness...constructs have been greatly streamlined; only two plasmids are required per system. Applications of these...sb500273n. Epub 2014 Oct 11. PubMed. Find the plasmids from this publication at Addgene. Synthetic biology...Chevalier AA, Tabor JJ, Simpson ZB, Lavery LA, Levy M, Davidson EA, Scouras A, Ellington AD, Marcotte EM, Voigt...2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication at Addgene. A synthetic...

  18. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...important collection.  Creating the Plasmids To begin constructing the plasmids for expression of mAbs, the Trimmer...expression plasmids were DNA sequenced to determine the VH and VL sequence. These plasmids have been deposited...NABOR are recombinant antibodies expressed from plasmids that have been deposited with Addgene. Here, we... first antibodies distributed by Addgene.  The plasmids used to generate these antibodies were based off...recombinant monoclonal antibody (R-mAb) expression plasmids, allowing for important reagents that could have...construction and validation of R-mAb expressing plasmids could be used by other labs to preserve and share...Addgene blog:  Plasmid-Based Recombinant Antibodies  Plasmids 101: Secondary Nanobody Toolbox Antibodies 101...

  19. dTAG - You're it!

    Type
    Blog Post
    ...1). Plasmids: We generated a series of plasmids that are available from Addgene. These plasmids enable...Resources on Addgene.org Browse all CRISPR plasmids CRISPR plasmids for protein tagging Find validated gRNAs...Detailed protocols describing the use of these plasmids are available here. The Methods section of Nabet...cloning procedures used to construct the deposited plasmids. Molecules: We synthesized dTAG molecules by conjugating...terminus should be tagged, the dTAG lentiviral plasmids can serve as a rapid means of comparing functionality...Cas9 Nickase Design and Homology Directed Repair Plasmids for endogenous gene tagging in human cells Resources...

  20. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...larger plasmids have lower rates of transfection. Minicircles: Minicircles are like mini-plasmids. They...choose a strategy suited to their research. Find Plasmids for Stem Cell Research Delivery Method Mechanism...expression of reprogramming factors. Non-replicating Plasmids: Generating iPSCs with plasmid-based expression...expression requires serial transfection of 1 or 2 plasmids that express the reprogramming factors of interest...express for longer periods of time than traditional plasmids due to lower activation of DNA-silencing mechanisms...minicircles are typically smaller than traditional plasmids, Jia et al generated iPSCs using a ~14.5kb minicircle...nuclear antigen-1 (EBNA1) based episomes: These plasmids carry the origin of replication (oriP) element...
Showing: 201 - 220 of 878 results