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We narrowed to 166 results for: crispr c plasmids

Showing: 141 - 160 of 166 results

  1. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    ...in the Lab Plasmids 101: Screens vs. Selections Plasmids 101: Knockout /Knock-In Plasmids ...the Lab Bench Plasmids 101 e-book Additional resources on the Addgene blog Plasmids 101: Using Transposons..., they eliminate the expression of the gene (by CRISPR knockout, shRNA depletion, etc.) and ask what the.... DOI: https://doi.org/10.1080/rsob.180241 Vanni, C., et al., Unifying the known and unknown microbial...

  2. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C., & Telesnitsky, A. (2008). Pseudodiploid Genome...Addgene's Lentiviral Guide Most popular lentiviral plasmids γ-Retrovirus Guide References Cullen, B. R. (...Vectors for Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives. Viruses, 13...34372494. Prussin, A. J., Garcia, E. B., & Marr, L. C. (2015). Total Virus and Bacteria Concentrations in...Xiao, H., Conway, J. A., Hunter, E., & Kappes, J. C. (1997). Functional RT and IN incorporated into HIV...

  3. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7 (2013): 1479-1491. PubMed...protein fusions Browse the Fluorescent Proteins Plasmid Kits ...Only about 10 amino acids can be deleted from the C-terminus and about 5 from the N-terminus, but this... standard practice to attach the GFP to the N- or C-terminus of another protein. This may, however, interfere...consensus sequence (MG-) and prenylation requires a C-terminal consensus sequence (-CaaX). Both modifications...when GFP is fused to these sequences. When N- and C-terminal fusions are not tolerated the alternative...the reasons that this works well is that the N- and C-terminus of GFP itself are relatively close, thereby...

  4. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...Mutate Your Plasmids Resources on Addgene.org Find Plasmids from the Whitehead Lab Use CRISPR Pooled Libraries... Can I use my favorite plasmid with this protocol? Maybe. Not all plasmids will work for the one-pot...strands of the plasmid will be nicked during the first BbvCI digest and the whole plasmid will be degraded...those in the plasmid. Again, if they are in the opposite direction, both strands of the plasmid will be nicked...map of your plasmid. If the sites are in opposite directions, you just need to use a plasmid with BbvCI...Generate Specific Mutations in Your Plasmids Learn How You Can Use CRISPR Pooled Libraries to Generate Mutant.... This technique relies on the generation of a plasmid library that expresses all desired variants of ...

  5. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ...a given disease. CRISPR has made genomic SDM relatively straightforward, but plasmid-based SDM has lagged...recommended for plasmids up to 25 kb, most cloners should be fine, especially using plasmids in the 10-12...Other Plasmid Cloning Techniques like SLIC and Golden Gate Cloning Learn How to Verify Your Plasmid Read...Reference Pages Choose Your Perfect Plasmid Backbone Find Plasmids for Your Research ...circular plasmids - no nick repair is needed. Since the bacteria are subsequently grown at 37 °C, the recombineering... REPLACR (Recombineering of Ends of Linearized Plasmids After PCR). Current site directed mutagenesis...mutation, then use those to PCR amplify the template plasmid. The parental, nonmutated DNA is digested with ...

  6. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...organism.   It’s not an overstatement to say that CRISPR/Cas9 editing is one of the key tools now providing...plans and in particular, that they all share the plasmids created under this programs via Addgene. New organisms...clutch of eggs? Propagation is thus quite difficult. CRISPR genome editing in squid is possible nonetheless...reptiles at all. But new success is coming with CRISPR/Cas in lizards. The Menke Lab was able to make ...sexual reproductive life cycles. One can inject plasmid DNA in the eggs to get transgenic animals. They...habitats - even 50 meters from Old Faithful in 60 °C soils. Dendrobatidae (Lauren O’Connell)  Poison frogs...

  7. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ...system. Currently, AAVs are being used to deliver CRISPR/Cas9 but have the potential to deliver other therapeutic...be used with new exciting technologies (such as CRISPR/Cas9) as they allow for gene delivery in an efficient...Gene Expression in Mammals A Match Made in Heaven: CRISPR/Cas9 and AAV Viral Vector Elements Additional ...Ready-to-Use AAV Preps Browse All of Our Viral Plasmids and Resources ...nitrogen or a dry ice/ethanol bath, and store at -80° C. Once thawed, the AAV aliquot should be used quickly...PubMed Central PMCID: PMC190186. 9. Grieger, Joshua C., and Richard J. Samulski. "Packaging capacity of ...

  8. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

    Type
    Blog Post
    Published
    Feb. 13, 2018, 2:51 p.m.
    ... protocols Browse CRISPR plasmids for isolating a genomic locus Browse FRET plasmids for studying protein...released from nuclei by a short incubation at 37 °C. These short DNA fragments can then be purified for...

  9. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...Biddle, M. S., Alende, C., Fotouhi, M., Jones, C., Ayoubi, R., Southern, K., Laflamme, C., Virk, H., NeuroSGC...T. M., Brown, C., Bandrowski, A., Virk, H., Edwards, A. M., McPherson, P., & Laflamme, C. (2023). Scaling....154034.1 Laflamme, C., McKeever, P. M., Kumar, R., Schwartz, J., Kolahdouzan, M., Chen, C. X., You, Z., Benaliouad...expressed at endogenous levels (Laflamme et al., 2019). CRISPR-Cas9 approaches are commonly used to produce KO... (tagged or untagged) protein When expression plasmids are available, or the necessary expertise to clone...involves the transient transfection of expression plasmids in a given cell line. For flow cytometry, it is..., H. S., Biddle, M. S., Smallwood, D. T., Weston, C. A., Castells, E., Bowman, V. W., McCarthy, J., Amrani...

  10. How to Write a Scientific Review Article

    Type
    Blog Post
    Published
    Feb. 16, 2017, 3:30 p.m.
    ...Hot Plasmids Articles to See Compiled Info on New Plasmids Review the CRISPR Field with Our CRISPR Guide...perfect. Good luck! References 1. Haery, Leila, Ryan C. Thompson, and Thomas D. Gilmore. "Histone acetyltransferases...

  11. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...research? Check out Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0...1.0, 1.1, 2.1 plasmids Brainbow 3.0, 3.1, 3.2 plasmids References: Transgenic strategies for combinatorial...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation... of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need a quick primer on how...Sato T, van Es JH, van den Born M, Kroon-Veenboer C, Barker N, Klein AM, van Rheenen J, Simons BD, Clevers...melanogaster. Hadjieconomou D, Rotkopf S, Alexandre C, Bell DM, Dickson BJ, Salecker I. Nature Methods. ...

  12. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...Fluorescent protein plasmids Use FRET to study protein-protein interactions Deposit plasmids with Addgene! ...Tagged protein can easily be overexpressed from a plasmid but overexpression may interfere with the system...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even...ti2learning.org. References 1. Dieterich, Daniela C., et al. "In situ visualization and dynamics of newly...

  13. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    Published
    July 27, 2017, 1:11 p.m.
    ...Photosactivatable Fluorescent Proteins Use Light to Control CRISPR Genome Editing Choose the Brightest Fluorescent...Addgene.org Find Fluorescent Protein Biosensors Browse Plasmids from the Tsien Lab Browse the Michael Davidson...light. Previous iterations of FLiPs only used N- and C-terminal dronpa fusions. With these various designs...

  14. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...all applications. Addgene’s plasmid collection includes over 10,000 plasmids with a FLAG tag (!), but did...cells By Susanna Stroik Even with the advances in CRISPR technology over the past decade, primary cells ... al. 2023 with permission.   Find Cas-PAGE plasmids here!   Zhang, Z., et al. (2023). Efficient engineering...inspired and informed this section.  Find mStayGold plasmids at Addgene here!   Ando, R., et al. (2023). StayGold...under CC-BY license.   Find the AAV.CAP-Mac plasmid here!   Chuapoco, M.R., Flytzanis, N.C., Goeden...(2023) under CC-BY license.   Find AAV1-X1 plasmid here! Chen, X., et al. (2023). Functional gene ...

  15. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes...crossover event between the vector plasmid and the packaging plasmid caused the Psi sequence to be placed...expression of the transgene   Transfer plasmids The transfer plasmid is the vector you clone your gene of...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection... additional protocols and plasmids.  References 1. Miller, A. DUSTY, and C. A. R. O. L. Buttimore. "Redesign...vectors fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral...techniques are still used and efficiently transfer plasmid DNA efficiently into dividing immortalized cells...

  16. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...down into 5 key steps: Construction of the rAdV plasmid (~1 week): The AdEasyTM system is the most popular...adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the transgene of interest...how to generate the final recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide...2.*Pro-Tip*: once the correct recombinant pAdV plasmid is identified, re-transform into a bacterial strain...recombination for amplification. The final purified plasmid stock should be analyzed by endonuclease restriction...-S). HEK293 cells are transfected with the AdV plasmid construct from step #1 and allowed to stay in culture...- a primer for beginners Adenoviral delivery of CRISPR/Cas9 5 tips for troubleshooting viral transductions...

  17. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ... first ever CRISPR-based clinical trial in 18 lung cancer patients. This study used CRISPR gene editing...T-regs) into their microenvironment by producing C-C motif chemokine 22 (CCL22) and TGFβ. T-regs are suppressive... Blog Learn About Our Cancer Collection Pages Plasmid Grant to Aid Cervical Cancer Screening in Ghana...

  18. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...Unpublished or pre-publication plasmids": Unpublished plasmids or plasmids that will not be published are...and entering a plasmid name,plasmid type, and plasmid description/experimental purpose (right). On the...with your plasmids, and adding all the plasmids at once increases the likelihood that the plasmids are given...easy for you to share plasmids. To achieve this goal, we will archive any plasmids you've deposited with...Spreadsheet". Start your plasmid deposit There are two options to begin your plasmid deposit. Figure 2: Choose... Choose to submit plasmid(s) online or by using a spreadsheet. The plasmid information can be entered ...than 10 plasmids but we recommend using the spreadsheet option for deposits of 10 or more plasmids.   ...

  19. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ....org Find plant plasmids Visit our plant resource page Deposit arabidopsis plasmids with Addgene ...this method include: Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a...available (Weigel and Glazebrook, 2002). Find plant plasmid resources at Addgene  Choosing an expression system...options, but here are a couple of my favorites. Plasmids from the Tsuyoshi Nakagawa lab are binary gateway...prefer Golden Gate assembly, there are a series of plasmids from the Dinneny lab that additionally allow for...culture you are growing definitely contains your plasmid. Grow lots of Agrobacterium: The more agrobacterium...Addgene blog Engineering the plant genome using CRISPR/Cas9 Using pSiM24 for simplifying plant genetic...

  20. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ...after transfecting with the transfer and packaging plasmids. A benefit of the multiple harvest strategy is...Elements Learn How to Do Genome-Wide Screens with CRISPR Lentiviral Vectors Additional Resources on Addgene.org... for less than a day, lentivirus can be kept at 4°C. For long-term storage, viral preps should be divided...divided into single-use aliquots, and stored at -80°C. Some reports suggest rapidly freezing virus in a ...
Showing: 141 - 160 of 166 results