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We narrowed to 217 results for: crispr cas9 expression plasmids

Showing: 121 - 140 of 217 results

  1. Hot Plasmids: Spring 2025

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    Blog Post
    ...empty and EGFP retroviral plasmid set, our bacterial plasmid set for expressing tagged proteins, and stay...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. ...backbones special: Strong expression: A CMV promoter drives robust gene expression in mammalian cells. Flexible...stay tuned for mammalian expression and AAV plasmid sets launching later this year!  Find Addgene's own...RNA-guided genome editing, think beyond Cas9 with STITCHR: a combined CRISPR / retrotransposon system with the...) Fluorescence from axonal boutons of neurons expressing iGluSnFR3, iGluSnFR4f, and iGluSnFR4s. C) Spike-triggered... synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A., Negrean, A., Chen, Y., Iyer...

  2. Plasmids 101: Golden Gate Cloning

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    Blog Post
    ... create a single gRNA-expressing plasmid, but it can also be adapted to express multiple gRNAs. Addgene...Golden Gate method to create CRISPR/Cas9 constructs, assemble standard plasmids parts in different combinations...specifying a gRNA target sequence into a Cas9-containing plasmid such as pX330. This cloning strategy not...Check out more Plasmids 101 Posts! Check out Addgene’s restriction cloning guide! CRISPR 101: Multiplex...Addgene’s plasmids are used with a wide variety of restriction enzyme-based cloning methods. Each method...Entry DNA overhangs may be present in the original plasmid (Option 1) or added using PCR-based amplification...fragments to create TAL effector nucleases. These plasmids were designed to utilize the BsaI and BsmBI Type...

  3. CRISPR 101: Drug discovery and CRISPR

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    Blog Post
    ...on the Addgene blog CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of...first time CRISPR User (by a first time CRISPR User) Additional resources on Addgene.org CRISPR Guide ... CRISPR Plasmids and Resources CRISPR History and Development for Genome Engineering    ...human trial phases.   So how does CRISPR factor into this workflow? CRISPR both speeds up the process and...potential targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens can be used...implementation of expression systems to accurately mimic a disease phenotype. CRISPR can also be used ...where CRISPR can save time and money to make sure quality candidates are selected.      CRISPR aids...

  4. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ...Editing CRISPR 101: Cas9 Structural Biology Additional resources on Addgene.org Browse CRISPR plasmids...how CRISPR-derived base editing works in general. Base editors use either catalytically dead Cas9 (dCas9...need from Cas9 are sequence specificity and the R-loop structure. In order to bind DNA, Cas9 requires ...to the stretch of ssDNA created by Cas9 binding. Pro tip! In CRISPR, the non-target strand is the DNA ...Acronym Full name Notes CRISPR terms gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a matching DNA...non-target strand of DNA for Cas9 to bind.   R-loop The structure formed by Cas9 binding to DNA and separating... Programmable C:G to G:C genome editing with CRISPR-Cas9-directed base excision repair proteins. Nature...

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

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    Blog Post
    ...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...about a recent plasmid deposit please sign up here.        A retroviral mouse CRISPR knockout library... 2 (Th2) cells from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation...using expression of differentiation factors as the screen readout. This retroviral mouse CRISPR pooled...dependent promoter systems. Find these CRISPR activation plasmids at Addgene. Liu et al., Nature Communications...library Article contributed by Alycia Bittner CRISPR pooled libraries are popular tools for genetic screens...

  6. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

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    Blog Post
    ...time CRISPR user? Check out our tips for 1st time CRISPR users Learn how to Choose the Best Cas9 Variant...biotin ligases Expression system Stable cell line expressing 1) FLAG-biotin-tagged dCas9, 2) BirA, and ...Find Other Cas9 Variants for Purifying Genomic Regions of Interest Here Brush Up on Your CRISPR Background...components are co-expressed, the gRNA targets the dCas9 to the loci of interest and dCas9 is biotinylated...the resolution, the Xu lab created a dCas9-based CAPTURE (CRISPR Affinity Purification in situ of Regulatory...creation of a stable cell line that co-expresses three components: dCas9 with an added biotin acceptor site...cell lines will have varying levels of dCas9 and gRNA expression. Additionally, CAPTURE 3C-based sequencing...

  7. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ....) CRISPR enzyme (Cas9 [sp], Cas9 [other], Cas12a, RNA-targeting) CRISPR mechanism (KO, CRISPRa, CRISPRi...resources from Addgene Plasmids 101: Modular Cloning Addgene's CRISPR Guide Plasmids 101: Golden Gate Cloning...restriction digest, and then plasmid sequencing if you can. To quote Doench, “Whole plasmid sequencing is just ...correctly banded clones for plasmid sequencing and found 98% of the plasmids were perfectly correct. Because...pRDA_757 (Piggybac; constitutive expression) Mammalian Plasmid pRDA_791 (contains only orio and...the assembled plasmid. It shows you each component, with the ID of the component plasmid, component name...the component plasmids on the Addgene website. Step 4: Naming your assembled plasmid and generating...

  8. Viral Vectors 101: Viral Applications

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    Blog Post
    ...applications Gene expression control Researchers are always wanting to manipulate gene expression – turn it up... depending on the vector and virus. Viral CRISPR-Cas9 tools have been engineered to introduce frameshift...interest, turning expression off permanently. Viral vectors can also be used to express proteins. Viral...Gene Therapy with AAV Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells... the genome of the target cell or transiently expressed. It’s important to note that for all the applications...sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily...Viral expression cassettes typically contain a promoter (inducible or constitutive), your gene of interest...

  9. Custom CRISPR Screens & the Green Listed Software

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    Blog Post
    ...Learn More About CRISPR Find CRISPR Screening Libraries Browse All CRISPR Plasmids ... custom CRISPR screens Differentially expressed genes  As the costs for global gene expression analyses...world class CRISPR labs.  Check Out This Video to See How Green Listed Works at a Glance!  CRISPR knockout...transcriptomics data. Custom CRISPR screens designed to target differentially expressed genes can be the next...test which of the differentially expressed genes found in a gene expression dataset are causal for a phenotype...differentially expressed genes that are known drug targets, or all differentially expressed genes that code... activity and minimize off-target effects of CRISPR-Cas9." Nature biotechnology 34.2 (2016): 184-191. ...

  10. Hot Plasmids February 2024

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    Blog Post
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...facilitates endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in ...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 ...lines and primary cells. Although cell entry of a Cas9 ribonucleoprotein (RNP) was limited (likely due ...multiplexed guides. Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization...Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter...

  11. Hot Plasmids - May 2022

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    Blog Post
    ...The dCas9-SSAP system reports an efficiency of up to 20%, comparable to commonly used CRISPR/Cas9 methods...subset of the new plasmids and viral preps 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...protein can then be co-expressed in essentially any transgenic zebrafish line expressing a GFP-tagged version...display increased neuronal expression in the brain while having decreased expression in the liver after systemic...only AAV.CAP-B10 causes brain wide expression, with no liver expression, after retro-orbital sinus vein ...mutations. These plasmids contain catalytically inactive dCas9 and microbial SSAPs (dCas9-SSAP) which insert...

  12. Lentiviral Vector Uses and Overview

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    Blog Post
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated... make stable transgene-expressing or knockout cell lines. Many CRISPR plasmids are designed for lentiviral... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with... of Addgene’s most requested plasmids are used with lentiviral expression systems. If you’re new to the...Addgene Blog Your Lentiviral Plasmid FAQs Answered Learn How Lentiviral CRISPR Libraries Enable Genome-Scale...Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional Resources on Addgene.org Check out...use, including Addgene’s top requested plasmid in 2015: lentiCRISPR v2, used both for targeted modification...

  13. Important Considerations When Using AAVs

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    Blog Post
    ...Versatile Viral Tool for Gene Expression in Mammals A Match Made in Heaven: CRISPR/Cas9 and AAV Viral Vector Elements... have found that expression occurs quickly and results in long and stable expression (up to 9 months in...the onset of gene expression. However, it has yet to be determined what gene expression looks like at 3 .... Currently, AAVs are being used to deliver CRISPR/Cas9 but have the potential to deliver other therapeutic...used with new exciting technologies (such as CRISPR/Cas9) as they allow for gene delivery in an efficient...AAV2 can transduce many tissue types, it has low expression in several brain regions where other serotypes... under study. Many AAVs will show significant expression in a variety of cell types, but you should try...

  14. 10 Great Guest Posts We're Thankful For

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    Blog Post
    ...you’re getting started with them. CRISPR Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening...publication. Optimizing Donor DNA for Enhanced CRISPR Genome Editing CRISPR has revolutionized how researchers make... an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “CRISPR-OS” In this post, Albert...Nathaniel Roquet from the Lu Lab at MIT developed plasmid based systems that report the order in which specific... action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (AAVs) are... can use CombiGEM to barcode small libraries of CRISPR gRNAs to use for screening experiments. Get the...and other types of genome edits, but, even with CRISPR, it isn’t always easy to insert a section of DNA...

  15. Plasmids 101: Screens vs. Selections

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    Blog Post
    ... Addgene Blog CRISPR libraries Genome-wide screens with CRISPR Lentiviral CRISPR libraries ....S. Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166): 80-84 (2014). doi...with small molecules/drugs. With the advent of CRISPR, genome-wide knockout screens are possible with...While the entire genome can be knocked out with CRISPR, the entirety of the small molecule realm is typically...downstream manipulations as a forward or reverse screen (CRISPR, drug, etc.), just with an added baseline component...component. Alternative screening techniques While CRISPR and drugs dominate the screening scene, there are... worth considering. RNAi screens are similar to CRISPR, but instead of knocking out the genes of interest...

  16. Antibodies 101: Validation

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    Blog Post
    ...include using RNAi to knock-down expression of your target or CRISPR/Cas9 to create mutations that essentially...invariable expression and non-specific, background signal, so you’ll need a range of expression strengths...gene for your protein has been mutated using CRISPR/Cas9. You check the mutation and it causes a large...that do not express your protein of interest endogenously and transfect them with a plasmid encoding your...Orthogonal Strategies This approach compares target expression of antibody-independent approaches to your antibody-dependent...proteins. Additionally, they require variable expression of your target across samples - it is hard to...neighborhood bioinformatician) to look at RNA expression of your gene of interest across a set of human...

  17. Zhang Lab CRISPR Page

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    Collection
    ...cloning information: CRISPR-Cas9 mouse toolbox protocol 637.8 KB CRISPR-Cas9 Cre expression vectors for cancer...PubMed . Return to AAV In vivo plasmids AAV Plasmids with Cas9 Mouse CRISPR-Cas9 knockin mice for genome editing...constitutively expressing Cas9 knockin mice (Platt et al ., Cell 2014). In these mice the CRISPR-Cas9 system ...sequence (20bp). CRISPR-Cas9 Cre expression vectors for genome editing in the brain Using Cas9 mice Platt et...pAAV-pMecp2-SpCas9-spA) for expressing Cas9 in neurons. SpCas9 guide acceptor: The PX552 plasmid (#60958)...Design Tools CRISPR Blog Posts Jump to: SpCas9 Plasmids GeCKO SAM AAV - In vivo Delivery AAV - Cas9 Mouse CRISPR...237.2 KB SpCas9 Plasmids The CRISPR/Cas system can be implemented in mammalian cells by co-expressing the ...

  18. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ...a 1st Time CRISPR User (by a 1st Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian...48-well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western ... Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...lines that overexpress genes of interest; more recently we have also been using Addgene CRISPR vectors to... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...and PCR, depending on the level of endogenous expression. We highly recommend the use of an electronic...ampules each of 4 clones with different levels of expression of our gene of interest. Advantages of the agarose...

  19. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

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    Blog Post
    ...these lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from a Tet/Shield1...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...write about a recent plasmid deposit please sign up here. Listen to the Hot Plasmids Podcast!  MarathonRT...at Yale University recently deposited a plasmid for expressing a novel reverse transcriptase (RT), MarathonRT...sequence of interest with an sgRNA expressed from a separate plasmid. When cells are treated with biotin–phenol...polymerase processivity. Please find the MarathonRT plasmid here! Zhao C, et al. RNA. 2018 PubMed PMID:29109157...

  20. Worm Expression Resources

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    Collection
    ... Plasmid Collections Worm Plasmids Worm Plasmids and Resources Plasmid Tools Worm Constructs... insertion using CRISPR/Cas9 and the selection of C. elegans strains. CRISPR plasmids from John Calarco's...elegans via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses purified Cas9, such as that... genes by feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily engineer specific...some of the plasmids and tools available at Addgene for use in worms. Browse Plasmids Plasmid Tools for ... Gene/Insert PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids that contain a worm...Addgene's CRISPR Guide for a general introduction to CRISPR technology. Addgene's worm CRISPR resources ...
Showing: 121 - 140 of 217 results