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We narrowed to 387 results for: plasmid dna

Showing: 121 - 160 of 387 results

  1. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    Published
    Aug. 16, 2018, 12:40 p.m.
    ...Split-BioID plasmids Find CRISPR plasmids for labeling genomic regions Find CRISPR plasmids for purifying...it feels like DNA and protein get all the attention.There are numerous ways to detect DNA-protein interactions...interest, much like using ChIP for studying protein and DNA interactions. Unfortunately, using proteins as bait... however, knock-in of BoxB sites into endogenous DNA sequences could allow one to study RNAs expressed...question that RaPID could help answer? The RaPID plasmids are available from Addgene! References Ramanathan...PubMed Central PMCID: PMC5886736. Find the RaPID Plasmids Here! McHugh, C.A., Russell, P., & Guttman, M...Additional Resources on the Addgene Blog Study Protein-DNA Interactions with CUT&RUN Study Protein-Protein Interactions...

  2. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    Published
    June 6, 2023, 1:15 p.m.
    ...synthesize complementary DNA copies of their genome upon infecting a host cell. This viral DNA (also called proviral...proviral DNA) is then inserted into the host's genome and is used to make more virus. The most studied ...their required components are very similar. While plasmids containing virus-specific elements such as the...Adeno-associated virus (AAV) is a small, single-stranded DNA parvovirus. AAV can infect humans and some other ...integrate, and the addition of a helper virus or plasmid allows it to replicate episomally, essentially ...can replicate and divide along with the endogenous DNA. AAV also has low immunogenicity, meaning it does...Adenovirus Adenovirus has a linear, double stranded DNA genome which is non-enveloped. It can infect humans...

  3. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...inducible inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al...designed the pOSIP (one-step integration plasmid) series of plasmids, vectors that convey both the sequence...recognition sites that flank a DNA module in an inverse orientation, so that multiple DNA components can directionally...mGRASP plasmids. Additionally, the lab recently described the use of another set of mGRASP plasmids in their... favorite plasmid technologies? Email us at [email protected]  and let us know what plasmids you would ...sgRNA libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA...

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

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...this evolving field! Find prime editing plasmids! Find plasmids from David Liu's lab! This post was originally...the CRISPR guide Find CRISPR plasmids by function Browse our entire plasmid collection ...possible without double-strand breaks (DSBs) or donor DNA templates.  Figure 1: An overview of prime...transcriptase. The Cas9 nickase is guided to the DNA target site by the pegRNA — a guide RNA that also...encodes the desired edit and homology to the genomic DNA locus. After nicking by Cas9, the homologous pegRNA...copies the desired edit, directly polymerizing new DNA onto the nicked target strand. The initial result... with overlapping strands of edited and unedited DNA. The heteroduplex is resolved by the cell’s mismatch...

  5. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    Published
    Dec. 12, 2013, 3:35 p.m.
    ...with the central dogma - replication of DNA, transcription of DNA to make RNA, translation of RNA to make...Drew Endy's lab at Stanford develops engineered DNA systems capable of storing data and computing inside...Addgene through the Biobrick Public Agreement (BPA) Plasmid Collection along with a BIOFAB kit comprised of..."Addgene is one of the leaders in distributing plasmids amongst the academic community. On the BioBricks... BioBricks Foundation to distribute the physical DNA samples, for when that's the preferred mode of receiving... with Endy more about the BIOFAB and BIL gates plasmid kits, the Biobricks Public Agreement, and the future...first to be made available via Addgene and the BPA Plasmid Collection - enable? Endy: The BIOFAB collection...

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

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...many cases several exons’ worth of DNA. Any ∼20 nucleotides of DNA can serve as the target for Cas9’s ...Prime editor engaging target DNA. The reverse transcriptase polymerizes DNA onto the target strand based...great choices. Check out our SpCas9 containing plasmids, optimized for a variety of species.   Specificity...Cytidine deamination takes place on the free strand of DNA and converts a C to U without producing a double ... platform to deliver various cargo to a specific DNA locus by directly fusing the cargo to dCas9.  This...frequently fused to Cas9 for this purpose. dCas9-KRAB plasmids for repressing target genes in a variety of species...additional RNA-binding transcriptional activator. Plasmids for activating target genes in a variety of species...

  7. Antibodies 101: ChIP

    Type
    Blog Post
    Published
    March 10, 2022, 3:25 p.m.
    ... treated so that the protein and DNA are no longer bound, and the DNA can then be isolated and studied...proteins of interest to any DNA they were bound to (2) fragmented the DNA (3) used antibodies to specifically...your antibody:protein:DNA complexes and finally (5) isolated the DNA from your antibody:protein:DNA complexes...preserve more of the protein:DNA complexes. Once you’ve finished fragmenting the DNA, spin down the samples...interactions, and can give valuable insight into DNA expression and protein interactions, including epigenetic...proteins are not likely to remain attached to the DNA throughout the sample processing and therefore will...modifying proteins, which bind quite strongly to DNA, crosslinking may not be necessary. This is known...

  8. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    Published
    June 16, 2015, 2:15 p.m.
    ... from Addgene Plasmids 101: How to Verify Your Plasmid Plasmids 101: Control Plasmids 6 Tips for Analyzing... on our plasmid pages - links to articles citing this plasmid. Now you can learn how a plasmid has been...your deposited plasmids Increased visibility of your plasmids in the "Associated Plasmids" lists Benefits...Find other related plasmids in the "Associated Plasmids" lists Properly citing plasmids you've requested...Citing This Plasmid" list. Email us at [email protected] with the Plasmid Name, Addgene Plasmid ID #, and...Troubleshooting DNA Sequencing Results ICYMI: Addgene’s Plasmids 101 eBook Protocols and Plasmid References...'ll find the plasmids listed below the citation. An example of these "Associated Plasmids" is shown in...

  9. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    Published
    April 23, 2020, 1:15 p.m.
    ...amount of spurious DNA from the packaging cell line, helper and packaging plasmids. We decided to use ... allows us to provide scientists with complete plasmid sequences. After launching our viral service in... the same frequency, we reasoned that, following DNA extraction, the [+] and [-] strands might naturally...restriction enzyme based methods, we showed that the DNA extract is a heterogeneous mix of single-stranded...double-stranded species. While not all of the molecules in the DNA extract associate with complementary strands, enough...identity of AAV genomes and detect contaminating AAV DNA. In addition, we wrote an open-source custom Python...Moullier P, Léger A (2015) Advanced Characterization of DNA Molecules in rAAV Vector Preparations by Single-stranded...

  10. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ...sequence is a strict requirement for Cas9-mediated DNA cleavage. The need for more PAM sequences While...S. pyogenes Cas9 that were able to cleave target DNA sequences located upstream of either NGA or NGC PAM...increase the specificity of genome modifications at DNA targets adjacent to NGG PAM sequences when used in...VRER Cas9 variants are capable of cleaving genomic DNA in mammalian cells and zebrafish embryos, and they...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 ...

  11. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ...Agrobacterium contains a plasmid with a T-DNA (transfer-DNA) region. During infection this T-DNA region is transferred...transformation Read our Plasmids 101 blog posts Resources on Addgene.org Browse our Plant Plasmids and Resources...page Find synthetic biology plant plasmids Find CRISPR plant plasmids ... natural process by replacing the genes on the T-DNA region with our gene of interest or a reporter gene...brought into contact with the leaf tissue and the T-DNA region with the gene of interest is transferred into...localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different leaf...

  12. Typing CRISPR Systems

    Type
    Blog Post
    Published
    March 18, 2025, 1:15 p.m.
    ... target both RNA and DNA, though it is largely believed that the cleavage of DNA is the primary function... foreign DNA into the CRISPR array. Lastly, type IV systems are most often found on plasmids. The many...Kyrpides, N. C., Banfield, J. F., & Doudna, J. A. (2018). Programmed DNA destruction by miniature CRISPR-...related immune mechanisms that could target foreign DNA with unique properties. This led to a need to develop...recently that researchers began building optimal plasmids to harness the power of Class 1 systems. Class...overall. Type I systems degrade large sections of DNA thanks to the dual action power of Cas3. Cas3 functions...helicase and a nuclease, unwinding and cutting up DNA after recruitment by Cascade. As you might imagine...

  13. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...on the Addgene Blog Plasmids for endogenous gene tagging in human cells Plasmids 101: Protein Tags Fluorescent... how proteins interact with one another (or with DNA) to perform diverse biochemical tasks in the cell...from many pitfalls: they require construction of plasmid vectors and/or rely on cumbersome in vitro synthesis...-IN” bioinformatics tool for guide RNA and donor DNA design, and implemented our optimised protocols into...to cells: a guide RNA, Cas9 protein, and a donor DNA that contains the tag sequence flanked by homology...CRISPR experiments (in collaboration with Integrated DNA Technologies, USA). These synthetic RNAs can also...csRNP complex formation, a single-stranded donor DNA (ssODN) that contains the tag-encoding sequence and...

  14. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    Published
    Sept. 6, 2018, 12:02 p.m.
    ...chosen for both Cas9 and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-...CRISPR plasmids) are not expressed. Because this method does not require the delivery of foreign DNA, and...commonly performed by transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains...targets with similar efficiency as compared to plasmid-based expression of Cas9/gRNA and can be used for...fluorophore tagged dCas9 may require the use of plasmid or viral-mediated delivery. You’ve decided to...and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex Schier’s Lab...experimental systems that may not be amenable to plasmid based methods. One of the most common techniques...

  15. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    Published
    Sept. 3, 2019, 1:15 p.m.
    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        New toolkit... the Kanemaki lab has created a new toolkit of plasmids for the N- or C- terminal tagging of proteins ... be from 2-6 base pairs in length and follow the DNA sequence targeted by the Cas9 nuclease. The Streptococcus...previously difficult. Find SpCas9 and Target-AID-NG plasmids at Addgene! Nishimasu H et al., Science, 2018....datasets for research and innovation. Find MerMAIDs plasmids at Addgene Opperman J et al., Nature Communications...

  16. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ...Nathaniel Roquet from the Lu Lab at MIT developed plasmid based systems that report the order in which specific...technology that can be used to quickly barcode a set of DNA sequences. In this post, Alan Wong (formerly of the...and the associated publication. Optimizing Donor DNA for Enhanced CRISPR Genome Editing CRISPR has revolutionized...CRISPR, it isn’t always easy to insert a section of DNA into the genome of your target organism. In this ..., describes how you can better design your donor DNA template to achieve more efficient genome editing...direct a variety of customizable functions to your DNA sequence of interest. Potential applications of the... plasmid...

  17. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...efficiency of plasmid delivery to a cell. In contrast, TREE provides a read out for plasmid delivery and...Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464–471 . https://doi.... to CRISPR/Cas9 edits that rely on double strand DNA breaks. Due to this inefficiency it is crucial for...enzymes. At the same time Cas9n cleaves the opposing DNA strain which results in two DSBs that can be repaired...panel of eGFP reporters to quantify the on-target DNA editing efficiency of APOBEC-Cas9 editisome complexes...circumvents the need for DSBs in monitoring on-target DNA editing efficiency of APOBEC-Cas9 editisome complexes...after being targeted by a cytidine deaminase base DNA editor. Specifically the BFP mutant (BFPH66) contains...

  18. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ... single stranded DNA (ssDNA) oligo which usually work better than plasmids. For ssDNA oligo design, the...easiest way is to co-transfect several plasmid together, with each plasmid having a targeting spacer cloned ...for PCR and surveyor. Q15: I used DNA polymerase Takara Ex Taq™ DNA Polymerase for my genomic PCR, but...consider using ssDNA oligo as donor template, it's usually more efficient than plasmid donor. Typical ...200ng to 500ng of 293FT genomic DNA as template after performing gDNA purification and have tried conditions...articles (3/12/2014). With so many options for CRISPR plasmid tools and numerous experimental design decisions...with expression, though it is possible for the plasmid to still express without the G. Because the G is...

  19. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ...number of plasmids found in each cell, but also whether the plasmid gets integrated into the host DNA or is...types of yeast plasmids are defined below: Yeast Integrating plasmids (YIp): These plasmids lack an ORI...our first few Plasmids 101 posts, we focused mainly on the elements required for plasmid maintenance within...integration. Yeast Episomal plasmids (YEp): These are most similar to bacterial plasmids and are considered “...well-defined genetic system, and a highly versatile DNA transformation system.Unlike most other microorganisms...simple gene replacement/mutation. Yeast expression plasmids used in the lab typically contain all the necessary...how the regulation of bacterial ORIs determines plasmid copy number within the bacterial cell. Similarly...

  20. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...your expression plasmid from these strains is not recommended as these auxillary plasmids may be isolated...control such as the pLys plasmid, which suppresses basal T7 expression. The pLys plasmid contains a chloramphenicol...arabinose-inducible plasmid will not express in an IPTG induction strain for example, nor will a p15 plasmid be compatible... for More Plasmids 101 Articles?  Find them all here: http://blog.addgene.org/topic/plasmids-101   ...In a previous Plasmids 101 blog, we reviewed the salient features of several popular strains of E. coli...coli for DNA propagation. While great for cloning purposes, these E. coli strains are not usually well...strain? Protein expression from high-copy number plasmids and powerful promoters will greatly exceed that...

  21. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...achieved at the level of the DNA construct, where your RNAi-target is fused to the DNA backbone that contains...for mammalian RNAi Browse all plasmids for RNAi Check out Addgene's plasmids and resources for plant biology...biological function in a direct or unbiased way at the DNA, RNA, or protein level. The “correct” choice of tool...Therefore, techniques that function downstream of DNA, such as RNA interference, can reversibly exert their...predominantly used to disrupt function at the level of DNA, thereby limiting CRISPR’s effectiveness at high ...initial considerations, such as the starting material (DNA-based vector or purified RNA) and the initial state...typically hairpin RNA, or artificial microRNA). Using a DNA template for the RNAi trigger is the most prevalent...

  22. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Alicyclobacillus acidoterrestris (AacCas12b) exhibits optimal DNA cleavage at 48 °C, a temperature incompatible to ...at 37 °C but preferentially nicks the non-target DNA strand. The lab then found gain-of-function mutations...paper in Nature Communications Find the Cas12b plasmids  Plant base editing with a Cas9-adenosine deaminase...wheat. Read the paper in Genome Biology Find plasmids containing these base editors Spatially and ...Neuroscience Find the AAV-based SaCas9 genome editing plasmids String assembly gRNA cloning (STAgR) String ...

  23. CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping

    Type
    Blog Post
    Published
    Nov. 29, 2018, 1:53 p.m.
    ...regions containing the putative tracrRNA into a plasmid and expressed it in an E. coli bacteria strain....References 1. Harrington, Lucas B., et al. "Programmed DNA destruction by miniature CRISPR-Cas14 enzymes." Science...sheet Resources on Addgene.org Find the Cas14 plasmids here Checkout our CRISPR guide page Browse the...indiscriminately cleave ssDNA in trans like Cas13 and Cas12 do for RNA and dsDNA, respectively. However...activated. Cas14 in diagnostics By cutting ssDNA but not dsDNA Cas14 is not a good candidate for a new genome-editing...combination enabling the detection of RNA, dsDNA, and now ssDNA. By comparing the ability of Cas12 and Cas14...and maybe more efficient Cas systems, Jennifer Doudna’s lab analyzed metagenomic datasets to try to determine...

  24. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ... in E. coli with two plasmids (Figure 3 below): Input Plasmid: The input plasmid (high copy, ColE1 origin...inducible promoter (PPhlF).  Output Plasmid: The output plasmid (single copy, bacterial artificial chromosome...through Addgene. Their plasmid maps and GenBank files can be found on their plasmid pages. All two-input...indicated figures in our paper (1)) Input Plasmid Output Plasmid Fig. 3A pNR64 pNR160 Fig. 4A pNR220...state machines (RSMs), which take on a particular DNA-sequence configuration (state) based on the identity...defined by the sequence within a prescribed region of DNA termed the “register”. Each chemical input to an ...result of their recombination is the excision of the DNA between the sites (Fig. 1B below). Alternatively,...

  25. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    Published
    April 16, 2020, 1 p.m.
    ... the Doudna lab has used Cas12a’s non-specific ssDNA degradation to develop the method termed DNA Endonuclease...specific DNA sequence, such as the Human papilloma virus (HPV) genome, via a crRNA. An ssDNA-fluorescently...signal when the ssDNA is degraded, is added to the reaction. To enhance sensitivity, the DNA is first amplified...quantified to determine the presence and quantity of DNA, RNA or a mutation of interest. Together SHERLOCK... increase the sensitivity of the assay, targeted DNA or RNA from a sample is first amplified using RPA...coupled with T7 transcription to convert amplified DNA to RNA for subsequent detection by Cas13. This amplification...with the ssRNA reporter enables SHERLOCK to detect DNA or RNA with atomolar sensitivity and single base ...

  26. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ...rescue! Base editors chemically modify DNA bases without breaking the DNA backbone, sidestepping the problem...uracil without cutting DNA. Uracil is then subsequently converted to thymine through DNA replication or repair...unmodified DNA strand so that it appears “newly synthesized” to the cell. Thus, the cell repairs the DNA using...strand of DNA and converts a C to U without producing a double strand break. UGI = uracil DNA glycosylase...Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature, 551(7681), 464–471. https...editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature, 533(7603), 420–424...homology-directed repair (HDR), the pathway for resolving DNA double-strand breaks (DSBs) preferred by researchers...

  27. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...incorporated into the DNA during DNA synthesis and then diluted with subsequent rounds of DNA synthesis (cell... for panel design. Find streptavidin plasmids and biotin plasmids at Addgene!     Figure 1: Indirect...found within the cytosol/nucleus, or visualizing DNA instead of proteins. Some stains can diffuse through...targets include transcription factors, histones, and DNA repair enzymes. Note that when using the nuclear ...Dye stains are non-antibody-based stains that bind DNA or free amines of proteins. Depending on the application...previously induced gene knockout. Besides amine- and DNA-binding dyes, nucleoside analogs like bromodeoxyuridine...stains are membrane-permeant or -impermeant and bind DNA or free amines, depending on the application. Paul...

  28. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...PubMed PMID: 26950006 Plasmid ID Plasmid Name Plasmid Type 74309 pcDNA3.0-TRPV4-p2A-ferritin-p2A-mCherry...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...features and applications of a partiular plasmid or set of plasmids. We hope that these articles make it ... you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below. With over...over 45,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...alone involves much literature searching, DNA synthesis, and plasmid assembly. Luckily, researchers from Douglas...

  29. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    Published
    Oct. 19, 2023, 1:15 p.m.
    ... ITRs are the elements that make an AAV plasmid an AAV plasmid.  Within the AAV genome, ITRs serve as...they are in the same DNA molecule as the gene that they package. In a transfer plasmid, the transgene is ...the single stranded DNA genome of AAV in the native virus. In an AAV vector plasmid, they flank the gene...growth culture and result in a high percentage of plasmid DNA with mutated ITR sequences. Using recombinant-deficient...gene of interest in a cell or animal model. AAV plasmids contain several important elements, but among ...located immediately outside of the ITRs in the plasmid backbone and do not properly assemble, but these...

  30. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids Topics: CRISPR, Other CRISPR... where we highlight a few of the newest CRISPR plasmids available at Addgene. If you want to read more...will cover: Miniature CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas...scaffold gRNA and making strategic mutations in the DNA binding cavity of Cas12f. The CasMINI system is less...Read the paper in Molecular Cell Find the CasMINI plasmids Efficient CRISPR editing with a hypercompact ...paper in Nature Biotechnology Find the Cas12f-GE plasmids Programmed genome editing by a miniature CRISPR-Cas12f...cells using various delivery methods, including plasmid, ribonucleoprotein, and AAV.  Read the paper in...

  31. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    Published
    Nov. 12, 2020, 2:15 p.m.
    ... usage bias in Plasmids 101: Codon usage bias Read more about plasmids in our Plasmids 101 blog series...without altering the sequence of the protein. As DNA synthesis has become quite cost-effective it seems...seems easy for scientists to re-synthesize DNA with the most optimized sequence. Unfortunately, it is not...algorithms/resources (Athey et al., 2017) and many DNA synthesis companies, such as IDT, Genewiz and GenScript...available online to both determine and synthesize DNA sequences for optimal expression in your system of... (2000) Codon usage tabulated from international DNA sequence databases: status for the year 2000. Nucleic...Reference Read about genetic code expansion Browse our plasmid collection page  ...

  32. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...transforming both fusion plasmids, or mating two single haploid yeast fusion plasmid transformants into a ... in the cell, including protein-protein, protein-DNA and protein-RNA interactions. The 1980s saw a flurry...format of some transcriptional activators: that the DNA binding domain (DBD) and transcriptional activation...reporter system (e.g. the Gateway-compatible bait Y2H plasmid pEZY202  from Yu-Zhu Zhang’s lab) is an example...interaction detection methods (6). In order to bind DNA and activate a reporter gene in eukaryotic cells,...Keegan, L., Gill, G., and Ptashne, M. Separation of DNA binding from the transcription-activating function...eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor. Cell. 1985...

  33. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...you will also need to deliver a donor plasmid or single-stranded DNA oligonucleotide, so you will need to...editing? Do you want to deliver the components as DNA, or would mRNA or protein delivery be more suitable...transduction or injection as either protein, RNA, or DNA. Finally, once you have identified the best expression...generate stable cell lines. Packaging and Envelope plasmids provide the necessary components to make lentiviral...for in vivo viral delivery. Cas9 mRNA and gRNA Plasmids containing gRNA and Cas9 are used in in vitro ...efficiency of CRISPR delivery. There are a few plasmids at Addgene that have been published as CRISPR ...genome target sequence into EGFP reporter. Target DNA reporter system: Can evaluate component delivery ...

  34. Twenty Years of Sharing Science!

    Type
    Blog Post
    Published
    Jan. 8, 2024, 3:25 p.m.
    ...repository for DNA-based materials, including our popular kits and libraries. This DNA-based materials...trying.  When we started in 2004, we were the plasmid repository. In 2016, we added viral vectors, and...addgene.org! Fun fact! Did you know you can search for plasmids from a publication by pasting the paper's title...Our first request was on September 8, 2004 (for pcDNA-p160MBP, if you're curious.) Since then, we've distributed...

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

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...Episomal vectors- extrachromosomally maintained DNA plasmids that express reprogramming factors To date,...viral origin of DNA replication, OriP and the EBNA-1 sequence that encodes for the DNA binding protein...and to model diseases in vitro. Find stem cell plasmids at Addgene Many sources of adult somatic cells... amplification of the episomes concurrently with DNA amplification in the host cell. This enables relatively...group is an optimized mixture of three episomal plasmids (Table 1) encoding the reprogramming factors. ...enhance the reprogramming efficiency, an extra plasmid, pCXWB-EBNA-1 is used in the reprogramming cocktail...increases protein expression from the other episomal plasmids. When used on PBMCs, the pCXLE episomal vector...

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

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...inhibiting crRNA loading, inhibiting DNA binding, or inhibiting DNA cleavage.   Five ways to anti-CRISPR...Bondy-Denomy J, Corn JE, Doudna JA (2017) Disabling Cas9 by an anti-CRISPR DNA mimic. Sci Adv 3:e1701620...proteins can also be used to prevent dCas9 tethering to DNA. Since these early anti-CRISPR studies, scientists...block CRISPR activity one of three ways: Inhibit DNA binding Example:  AcrIIA4 blocks Cas9’s interaction...crRNA-Cas complex (Zhu et al., 2019). Inhibiting DNA cleavage Example: AcrIIC1 binds to the HNH endonuclease...endonuclease domain of Cas9 and prevents target DNA cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR...CRISPR   Resources on Addgene.org Find CRISPR Plasmids for Your Research Interested in light-inducible...

  37. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...Read blog posts related to plasmid protocols Find protocols for purifying DNA without a kit Resources ...precipitate RNA, but not DNA and proteins. Lastly, using a DNase (there are several DNase enzyme products on...As with DNA isolation, scientists commonly rely on RNA isolation kits to make their life easier. Recently...Recently, we published a blog on DNA purification without a kit that outlined several reasons why doing ... one should take care whenever doing any type of DNA or RNA purification to avoid contamination, take ...RNA extraction. RNA is inherently not as stable as DNA- it is single stranded and its ribose groups are ...There are many alternative methods for isolating DNA without a kit. However, that isn’t the case for RNA...

  38. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...its method of DNA cleavage. Instead of cleaving DNA at a defined site to form a clean DNA break, Cas3 locates...sequence and initiates single stranded DNA degradation. Cas3 DNA digestion can proceed up to multiple kilobases...fidelity ssDNA targeting. RNA targeting Cas enzymes Move over Cas9, CRISPR isn’t just for DNA anymore!...highlighting their similarities and differences to Cas9. DNA targeting Cas enzymes Cas9 – The genome engineering...genome where it will proceed to generate a blunt DNA double strand break. Cas9 has been studied for genome...expanding number of others. Best for: a wide range of DNA edits.   Cas3 – The shredder What sets Cas3 apart...but it is effective at wiping out large regions of DNA and has anti-viral application potential as a result...

  39. What's New in CRISPR - May 2019

    Type
    Blog Post
    Published
    May 28, 2019, 12:49 p.m.
    ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...genome-wide CRISPR inhibition Covalent tethering of DNA template to Cas9 SECURE base editors Nme2Cas9 CRISPR...CRISPR inhibition library Covalent tethering of DNA repair template to Cas9 Homology-directed repair ...Communications Biology Find the Cas9-PCV and PCV-Cas9 plasmids Selective curbing of unwanted RNA editing (SECURE...paper in mSphere Find the dCas9 and dCas9-Mxi1 plasmids If you have a new CRISPR tool you’ve recently...

  40. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...editing of a target base in genomic DNA without double-stranded DNA cleavage." Nature 533.7603 (2016): ...Programmable base editing of A• T to G• C in genomic DNA without DNA cleavage." Nature 551.7681 (2017): 464. PubMed...all CRISPR plasmids and resources at Addgene Find CRISPR base editors Find CRISPR plasmids for plants ...frequencies and relies on the efficient delivery of donor DNA to facilitate repair. Since the development of CRISPR-mediated...Cas9 nickase (nCas9). nCas9 creates a nick in the DNA by cutting only a single strand, dramatically reducing... detrimental indel formation. After binding with DNA, the CBE deaminates a target cytosine (C) into a ...this change. However, there are no naturally known 'DNA adenosine deaminases' which would theoretically give...
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