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We narrowed to 284 results for: cas9 plasmid

Showing: 141 - 160 of 284 results

  1. 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 method..., relying on established reagents such as CRISPR/Cas9, Tol2, and the high-quality sequenced zebrafish ..., and their resistance (so far) to facile CRISPR/Cas9-mediated insertion of foreign DNA. Despite these...collection Read our molecular biology reference Find plasmids for your research ...

  2. 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...(Thomason et al., 2014). In recent years, CRISPR-Cas9 technology has also aided in the creation of scarless...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... an allelic exchange experiment? Find the pTOX plasmids here! Continued development of new allelic exchange...15109831. Pyne, Michael E., et al. "Coupling the CRISPR/Cas9 system with lambda red recombineering enables simplified...

  3. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...restriction enzyme or PCR to make linearized vector, the Cas9 enzyme along with a specific gRNA was used to cleave...Ligation Independent Cloning (SLIC) in a previous Plasmids 101 post. Although SLIC may be more cost effective...Li K, Wang B, Jin S, Reiser M, Lockey RF. CRISPR/Cas9 nuclease cleavage combined with Gibson assembly ... Zhao X, Gabrieli T, Lou C, Ebenstein Y, Zhu TF. Cas9-Assisted Targeting of CHromosome segments CATCH ...Resources on the Addgene Blog Browse All of Our Plasmids 101 Posts Read About Other Cloning Techniques ...Techniques Additional Resources on Addgene.org Find Plasmid Cloning Protocols Find Virus Protocols Find Basic...

  4. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    Blog Post
    ...following components: A plasmid in the host bacterium expressing dCas9 A plasmid in the host bacterium ...compatible with all Cas9 and Cas12 homologs that the researchers tested, including SpCas9, SaCas9, LbCas12a, enAsCas12a...the DNA. ABEs are somewhat slow and need a strong Cas9 domain to “hold on” long enough to the DNA to induce... of gene III to create infectious particles. A plasmid-based genetic circuit in the bacteria links the...encoding the deaminase TadA-7.10 A mutagenesis plasmid controlling the phage mutation rate Phage infection...Phage-assisted evolution of base-editing activity. A plasmid-based genetic circuit in the bacteria links the...off-target RNA editing rate of ABE7.10. Find the ABE8e plasmids at Addgene! Why engineer and optimize base editors...

  5. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...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... Mammalian Plasmid pRDA_791 (contains only orio and ampR) Drosophilia Plasmid pRDB_051 (for...depending on above selections.) CRISPR enzyme (Cas9 [sp], Cas9 [other], Cas12a, RNA-targeting) CRISPR mechanism...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...resources from Addgene Plasmids 101: Modular Cloning Addgene's CRISPR Guide Plasmids 101: Golden Gate Cloning...

  6. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...specifying a gRNA target sequence into a Cas9-containing plasmid such as pX330. This cloning strategy not...Gate method to create CRISPR/Cas9 constructs, assemble standard plasmids parts in different combinations...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...makes it easy to create a single gRNA-expressing plasmid, but it can also be adapted to express multiple...PubMed. More Resources at Addgene:  Check out more Plasmids 101 Posts! Check out Addgene’s restriction cloning...

  7. Hot Plasmids - October 2020

    Type
    Blog Post
    ...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...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head... Get the plasmids at Addgene Sc++ is a codon-optimized, engineered Streptococcus canis Cas9 nuclease ...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...head over to our CRISPR Plasmids and Resources page.  CasPhi is 70 kDa Cas protein identified exclusively...nuclease that has a broad 5′-NNG-3′ PAM. Get the plasmids at Addgene The Human GlycoGene CRISPR Library is ...

  8. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...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...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...head over to our CRISPR Plasmids and Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair...linked SaCas9 ABE variants. It's also one of the smallest AAV-deliverable ABE.  Find the plasmid here ...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the viral service A fast blue-light...

  9. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    ...example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids), but so are lentiviral vectors... deliver Cas9 and other genes to mammalian cells. For this reason, the top requested plasmid of 2017 is...envelope plasmid pMD2.G from Didier Trono’s lab! Lentivirus envelope vector pMD2.G This plasmid has been...Plasmid technologies are constantly evolving, but sometimes a technology is so useful it forever enhances...the broad usefulness of pMD2.G and VSV-G. This plasmid has been used in studies of the secretome, ubiquitin...to check out the Trono Lab website. If you have plasmids you’d like to deposit with Addgene, be sure to...Who knows, maybe yours will be the top requested plasmid of 2018! Additional Resources Quick Guide to ...

  10. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...Addgene.org Browse our CRISPR plasmids for visualization Find plasmids for your research Read our CRISPR...fluorescent protein to various Cas9 variants, SpCas9, NmCas9, and St1Cas9. These fusions localized to specific...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without..., give CRISPR-Sirius a try. Find CRISPR-Sirius plasmids at Addgene. References Ma, Hanhui et al. “Multicolor...inhibition, among others. Over the past few years, dCas9 has also become a robust visualization tool to study...University of Massachusetts Medical School developed a dCas9-based multicolor labeling system to visualize chromosomal... al. “Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.” Nature...

  11. It’s CRISPR Week at Addgene!

    Type
    Blog Post
    ...received the first CRISPR plasmid deposit. Now we have over 8,000 CRISPR plasmids in the repository that ...knockouts using CRISPR and Wednesday will highlight some Cas9 variants. Next, we’ll look at various CRISPR applications...

  12. Addgene’s Top 10 Blog Posts from 2014

    Type
    Blog Post
    ...Patrick CRISPR-Cas9 FAQs Answered! By Caroline LaManna Plasmids 101: What is a plasmid? By Margo R....loved helping to answer your plasmid and cloning questions through our Plasmids 101 series and by responding...Melina Fan Plasmids 101: The Promoter Region – Let's Go! By Kendall Morgan Plasmids 101: Mammalian...Joanne Kamens Plasmids 101: Antibiotic Resistance Genes By Marcy Patrick Plasmids 101: Yeast Vectors...as seen through the lens of Addgene’s blog and plasmid repository. Our blog is just over a year old, and...detailed information about this new technology. Our Plasmids 101 series continues to grab scientists’ attention...Genome Editing By John Doench (Guest Blogger) Plasmids 101: Origin of Replication By Kendall Morgan ...

  13. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ... our CRISPR guide Find CRISPR plasmids for your research Browse plasmids for base editing References ... the genome. They fused a catalytically impaired Cas9 protein to a DNA-editing enzyme to generate the ...based on a prime editor which is a fusion between Cas9 and reverse transcriptase. Using an elongated template...The new frontier of genome engineering with CRISPR-Cas9. Science 346:1258096 . https://doi.org/10.1126/science...Zhang F (2013) Genome engineering using the CRISPR-Cas9 system. Nature Protocols 8:2281–2308 . https://doi.org...

  14. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...single plasmid transformation, though, a pooled library has hundreds to millions of distinct plasmids, all...anyone who has worked with individual plasmids knows, some plasmids amplify well and others don’t. The size...pooled library, which contains a collection of plasmids in a single tube, or a pre-made viral prep library... prepare the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral...library and appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus...amplify the library. This is similar to amplifying a plasmid: transform E. coli with the pooled library DNA,...own protocol that amplifies without disrupting plasmid representation. A good place to start is with a...

  15. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... involving two-plasmid or single-plasmid systems (Vo et al., 2020). In the two-plasmid system, an effector...effectors employed by Type II systems (such as CRISPR-Cas9). Unlike in regular Type I CRISPR-Cas systems, however...group’s initial report of VchINT, three separate plasmids and multiple promoters were used to express the...effector plasmid (pEffector) expresses all seven required genes and the CRISPR RNA under the same promoter...promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also be combined...combined in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and ...and the ability to easily cure cells of the plasmid after integration. Figure 2: (A) Streamlined ...

  16. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...unique sgRNAs into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This allows for the multiplexing...repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized CRISPR ...between CRISPR components expressed from different plasmid backbones. They noted that backbone differences... as the order, or distance in the placement of plasmid elements can impact expression levels and consequently...system was used to assess how different lentiviral plasmid backbone elements impact viral titer and to compare...plants, and animals (Chiasson et al., 2019). Many plasmid elements such as promoters or fluorophores are ...High-Efficiency Genome Editing in Plants Mediated by a Cas9 Gene Containing Multiple Introns.” Plant Communications...

  17. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily...depending on the vector and virus. Viral CRISPR-Cas9 tools have been engineered to introduce frameshift...transfection and random genomic integration of a plasmid. Viral transduction is also generally less toxic... Therapy with AAV Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells ...

  18. CRISPR Cheat Sheet

    Type
    Blog Post
    ...RNA sequences. Cas9 is a very commonly used CRISPR nuclease. Note: Most CRISPR plasmids from Addgene ... to deliver. Not all cells efficiently take up plasmids used to produce CRISPR systems    ...versions of CRISPR nucleases that can’t cut DNA (dCas9, for example) that are used to increase or decrease...

  19. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...cargo via Cas9-directed integrases.   Use PASTE can be delivered via a single dose of plasmids, works...PASTEsystem involves insertion of landing sites via Cas9-directed reverse transcriptases, followed bylanding...adenovirus templates. The system, which is larger than Cas9, can also be used with mRNA and a synthetic guide...

  20. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ... and semen samples. Ma et al. designed plasmids containing Cas9 and two different gRNAs, and electroporated...genome sequencing in selected control and CRISPR-Cas9 injected embryos. In all cases, they did not observe...CRISPR Resources on Addgene.org Browse All CRISPR Plasmids Check Out Our CRISPR Guide Pages ...
Showing: 141 - 160 of 284 results