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We narrowed to 230 results for: crispr c plasmids

Showing: 121 - 140 of 230 results

  1. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...- to C-terminus. For instance, Lifeact-mTurquoise2 (Plasmid #36201) or mScarlet-tubulin (Plasmid #85045...fluorescent protein is at the C-terminus of the POI or that the POI is at the C-terminus of the fluorescent...revolution in genome editing methods (most notably CRISPR/Cas mediated HDR) enables the tagging of endogenous...advisable to first express the fusion protein from a plasmid-based system to assess whether the chimera is still...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...protein fusions Browse the fluorescent proteins plasmid kits ...dimerization (Shaner et al., 2008 - supplementary figure C). However, these assays are qualitative. The Organized...

  2. Top Requested Lentivirus and AAV of 2016

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    Blog Post
    ...lentiviral inventory is based on CRISPR tools, though we do provide a few non-CRISPR related viral preps. Cas9...Cas9 is the headliner of most CRISPR applications and can be expressed from your cell line of choice after...typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be used to generate ...can then be used both for screens (using pooled CRISPR knockout libraries) and individual gene knockouts...with a single-vector system). Tips for using this plasmid Cas9 expression is typically cytotoxic, which makes... Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4. PubMed...Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, Maratos-Flier E, Roth...

  3. CRISPR 101: Making a Knock-In Cell Line

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    Blog Post
    ...Efficiency for CRISPR Editing in Human Cells Resources on Addgene.org Browse All CRISPR Plasmids Find Validated...for successful HDR events. CRISPR cut site location Position your CRISPR cut site as close as possible...A., Fernandez, J. P., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1 mediates efficient homology-directed...experiment. Also learn about Addgene’s knock-in plasmids for reagents to help you knock your knock-in experiment...s41589-019-0432-1 Mehdi Banan. Recent advances in CRISPR/Cas9-mediated knock-ins in mammalian cells. J Biotechnol... homology-directed repair choice in response to CRISPR/Cas9 Induced-double strand breaks. Int J Mol Sci...human genome engineering by controlled timing of CRISPR/Cas9 delivery. eLIFE. (2014) 10.7554/eLife.04766...

  4. Five Popular Model Organisms

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    Blog Post
    ... eBook Read our plasmids 101 blog series Resources on Addgene.org Browse our plasmid collection pages... et al., 2015). If you are interested in C. elegans plasmids and resources check out Addgene’s Worm Expression...an ortholog in C. elegans making it ideal to study basic molecular biology processes. C. elegans are also...PMCID: PMC5525645. Mello, Craig C., et al. "Efficient gene transfer in C. elegans: extrachromosomal maintenance... resistant to chemotherapy. You can find these plasmids at Addgene. Another advantage of mice as model... Mice can be easily manipulated with tools like CRISPR to make transgenic lines. The fruit fly (Drosophila... pancreas. They first tagged these cells using plasmids. Then using HOTcre, a method that uses heat induction...

  5. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...of the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles. .... K., Chen, J., Pommier, G. C., Cogan, J. Z., Replogle, J. M., Adriaens, C., Ramadoss, G. N., Shi, Q.,...be used for applications beyond cryo-EM.   Find plasmids for MagIC-Cryo-EM here!   Arimura, Y., Konishi...disease phenotype in an animal model. Find PE-eVLP plasmids here! The lab’s other recent deposit builds on...over the wild-type enzyme. Find optimized PASSIGE plasmids here!     Figure 3: Schematic of the PASSIGE...recombinase integrates a sequence from a donor plasmid containing a complementary site into the genome...without compromising their efficacy.   Find CHARM plasmids here!   Neumann, E. N., Bertozzi, T. M., Wu, E...

  6. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries for CRISPR screening...heat-inactivated FBS and 5mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and FBS lots can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...dose of antibiotic, which may not be stable at 37 °C. To achieve a stable cell pool, the antibiotic selection...well plate and left undisturbed for 13 days. (a, b, c) Colonies formed by expansion of single cells for ...

  7. Educational Planarian Tools for Teaching Developmental Biology Techniques

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    Blog Post
    ...learning about CRISPR Resources at Addgene.org Find plasmids from the Sánchez lab Find plasmids for other...experiments in planarians with their recent plasmid deposit. These plasmids enable two main types of experiments... cloned into plasmids to investigate for each of these planarian species. These plasmids all have a PR-T4...insert the cDNA into the desired plasmid backbone. The Sánchez lab plasmids allow students and new labs to...Williams, Eric J. Ross, Sofia M. C. Robb, Sarah A. Elliott, Kimberly C. Tu, Alejandro Sánchez Alvarado,...of specific gene expression. The 49 deposited plasmids enable students to study gene expression and functions... dsRNA-induced RNAi experiments. Notably, this plasmid harbors a multi-cloning site flanked by T7 promoters...

  8. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...PMC3223083. 6. McInnes, J. L., Forster, A. C., Skingle, D. C., & Symons, R. H. “Preparation and uses of...PMC164612. 11. Liou, Y.-R., Wang, Y.-H., Lee, C.-Y., & Li, P.-C. “Buoyancy-Activated Cell Sorting Using Targeted...biotinylating proteins with BirA. You can find plasmids that contain both AviTag and BirA at Addgene! ... interactions Learn how biotin can be used with CRISPR Additional resources ThermoFisher Scientific...fusion in which AviTag is added to either the N or C terminus of the protein. BirA has been shown to effectively...

  9. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ... 25°C. At low temperatures like 18°C, flies take longer to develop. At high temperatures like 30°C, flies...Drosophila Plamids Drosophila CRISPR Resources Deposit Your Drosophila Plasmids ...have fly collections for in vivo work, including CRISPR tools. The Vienna Drosophila Resource Center ...

  10. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ...genome-wide CRISPR knockout screen to analyze the following unanswered questions about CRISPR screen design...genome-wide CRISPR/Cas9 screen design Are non-targeting guides an appropriate control? Many CRISPR screens...new approach to genome-wide CRISPR screen design.   Bassik lab’s CRISPR knockout libraries Overview...expected to be identified in a CRISPR knockout screen. Improving the Design of CRISPR Knockout Screens Before...conduct genome-wide screens with CRISPR/Cas9 Review the basics of CRISPR with this cheat sheet Read our ...Check out this guide to pooled plasmid libraries Find all of the CRISPR Pooled Libraries currently available...Genome-wide CRISPR/Cas9 screens are a high-throughput systematic approach for identifying genes involved...

  11. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    ...our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression...protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins at the beginning.... Now, we've turned some of the most requested plasmids into lentiviral preps to speed up COVID-19 research...now available as a lentiviral prep. It's a human CRISPR inhibition library that inhibits over 18,000 human...Testing AAV inventory for the entire collection! Plasmid Serotype Name Depositor 27056 AAVrg pAAV-Ef1a-DIO...constructs that allow for targeted expression. Plasmid 75470 is now available in AAV1 for Flp-dependent... mCherry reporter. We now also offer AAVrg for plasmid 83898, which targets expression of ChR2 to forebrain...

  12. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ...Delivering CRISPR Ribonucleoproteins Resources on Addgene.org Browse All CRISPR Plasmids Check Out Our...Increasing HDR accuracy with CRISPR/Cas-blocking mutations After genome editing with CRISPR/Cas9 in human cells...Paquet, et al  2017). Find Additional CRISPR Resources on Addgene's CRISPR Guide Page 2. Optimizing "cut-to-mutation...silent CRISPR/Cas9 blocking mutations in either the PAM (changing one of the G’s to an A,T or C – but ...s lab at the Rockefeller University in NYC. The CRISPR/Cas9 system is a versatile tool for precise gene... many organisms and model systems. We have used CRISPR/Cas9 extensively for the purpose of making sequence-specific...human induced pluripotent stem cells (iPSCs). The CRISPR/Cas9 com­plex is very efficient at introducing ...

  13. Plasmids 101: Modular Cloning Applications and Kits

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    Blog Post
    ...more of these kits, as there are many options! CRISPR CRISPR-Cas can harness the power of MoClo to expand...repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized CRISPR kits too... made comparisons between CRISPR components expressed from different plasmid backbones. They noted that...comparisons of CRISPR parts. The Fragmid system was used to assess how different lentiviral plasmid backbone...including conditional gene expression, multiplexed CRISPR, and chromosomal integration of larger expression...targetability and effectiveness. Perhaps the most intuitive CRISPR MoClo application is to assemble multiple unique...edit optimization, and more. While there are many CRISPR-Cas libraries and tools available for certain model...

  14. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...background via P1 transduction. Plasmid Expressing lambda red genes from a plasmid allows for a mobile recombineering...Repair Use Knockout/Knockin Plasmids to Disrupt Genes in Mammalian Cells CRISPR Methods for Bacterial Genome...MMR activation.  The first way is to introduce a C/C mismatch at or within 6 base pairs of the edit site...-50% when activation of MMR is avoided Introduce C/C mismatches or silent changes in wobble codons to ...recombination genes at low temperatures (30-34˚C).  Shifting bacteria to 42˚C for 15 minutes quickly inactivates ... flexibility to modify the E. coli chromosome, plasmid DNA or BAC DNA.  To use the lambda red recombineering...bacteria with integrated defective prophage 2) from a plasmid, 3) from mini-λ or 4) from the lambda red phage...

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

    Type
    Blog Post
    ...RNAs.   RNAi vs. CRISPR: Complementary techniques It is fair to say that CRISPR has revolutionized ...our CRISPR blog posts Resources on Addgene.org Find tools for mammalian RNAi Browse all plasmids for...unprecedented ways. With the meteoric rise and diversity of CRISPR technologies, one might incorrectly relegate RNAi...straightforwardness of RNAi is unmatched—unlike CRISPR experiments that require exogenous protein, successful...only requires the presence of dsRNA. In plants, CRISPR technologies are predominantly used to disrupt ...function at the level of DNA, thereby limiting CRISPR’s effectiveness at high copy numbers and genomic...prevalent assumption into question. Specifically, CRISPR-generated mutants believed to be knockouts can ...

  16. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria." Plasmid 51.3 (2004): 246-255. PubMed... on the Addgene blog Read our Plasmids 101 blog posts Find our CRISPR blog posts Learn more about Lambda...genomes using exogenous enzymes: Lambda red and CRISPR Developed over the past 20 years, Lambda Red recombineering...used  (Thomason et al., 2014). In recent years, CRISPR-Cas9 technology has also aided in the creation ...exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate off-target mutations...a mutated allele introduced from a conjugative plasmid replaces the chromosomal copy. This is a versatile... These two steps are (Figure 1): The initial plasmid integration step (the so-called “single crossover...

  17. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...Read our plasmids 101 blog series Resources on Addgene.org Browse our zebrafish CRISPR plasmid collection...zinc-finger nucleases, TALENs, and more recently CRISPR/Cas9. Another popular targeted gene deactivation...developed, relying on established reagents such as CRISPR/Cas9, Tol2, and the high-quality sequenced zebrafish...systems, and their resistance (so far) to facile CRISPR/Cas9-mediated insertion of foreign DNA. Despite...collection Read our molecular biology reference Find plasmids for your research ...PubMed Central PMCID: PMC6247916. Felker, A., and C. Mosimann. "Contemporary zebrafish transgenesis with...

  18. Worm Expression Resources

    Type
    Collection
    ... Plasmid Collections Worm Plasmids Worm Plasmids and Resources Plasmid Tools Worm... insertion using CRISPR/Cas9 and the selection of C. elegans strains. CRISPR plasmids from John Calarco's...Heritable genome editing in C. elegans via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses...some of the plasmids and tools available at Addgene for use in worms. Browse Plasmids Plasmid Tools for ...Addgene's CRISPR Guide for a general introduction to CRISPR technology. Addgene's worm CRISPR resources ... PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids that contain a worm ( C. elegans...Split cGAL plasmids - Paul Sternberg Lab. Plasmids for the temperature-robust GAL4-UAS system in C. elegans...

  19. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    ...interest has been cloned into an expression plasmid, the plasmid can be introduced into host cells, such ...that the plasmids and sequences for antibodies are not shared. For instance, through open plasmid sharing...Recombinant monoclonal antibodies are encoded in plasmids and produce a single antibody recognizing one ...synthetic genes that are typically expressed from a plasmid or from an integrated sequence in a stable cell...heavy chain genes of the antibody are encoded in a plasmid. (Right) An assembled antibody protein binds an...laboratories and companies have already begun creating plasmids encoding affinity reagents. These include conventional...performed will be useful in perpetuity.  Second, plasmids are easy to store and share, making recombinant...

  20. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...Resources on Addgene.org CRISPR Plasmids – Protein Tagging CRISPaint Gene Tagging Plasmids 101 eBook Resources...Resources on the Addgene blog Plasmids 101: Protein Tags Plasmids for Endogenous Gene Tagging in Human Cells...cloned in-frame to the protein of interest for plasmid-based expression systems. Degron tags are one of...several advantages over the alternative systems – CRISPR, siRNA, etc. Firstly, they are reversible mechanisms...protein tag which can be added to either the N or C terminal of a protein and can inducibly facilitate...
Showing: 121 - 140 of 230 results