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We narrowed to 208 results for: crispr cas9 expression plasmids

Showing: 161 - 180 of 208 results
  1. CRISPR 101: Any Base Transversion Editors

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    ...Addgene.org Browse CRISPR plasmids for base editing Get a broad overview with our CRISPR Guide Check out ...Resources on the Addgene Blog CRISPR 101: Cytosine and Adenine Base Editors CRISPR 101: Cytosine Transversion...activity. To drive selective A → T editing, they co-expressed Polη, a translesion synthesis polymerase known...inserted the deaminase into the C-terminal domain of Cas9, which narrowed the editing window and reduced A...site. Like in most other base editor designs, the Cas9 nicks the non-edited strand to encourage the cell...engineering going on in the base editing field! From CRISPR-free approaches to mitochondrial base editing to...out our CRISPR 101 ebook ...
  2. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    ...RNAs.   RNAi vs. CRISPR: Complementary techniques It is fair to say that CRISPR has revolutionized ...A, Bassik MC (2016) Systematic comparison of CRISPR/Cas9 and RNAi screens for essential genes. Nat Biotechnol...our CRISPR blog posts Resources on Addgene.org Find tools for mammalian RNAi Browse all plasmids for...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region...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 ...
  3. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

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    ...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...snail can be manipulated by injected RNA and can express fluorescent proteins. This lab has created an automated...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...-Mediated Ovary Transduction of Cargo)  delivers Cas9 RNP to the arthropod germline by injection into ...
  4. CRISPR Plasmids - Purify Genomic Loci

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    ...purification using CRISPR and other CRISPR topics, read our CRISPR Guide . Mammalian ID Plasmid Gene/Insert ...more CRISPR functions along with plasmids categorized by organism by visiting our CRISPR plasmids page...list of CRISPR resources by visiting our CRISPR reference page . CRISPR Guide Viral Preps CRISPR Protocols...Publication Return to top CRISPR Resources Addgene has a large selection of CRISPR plasmids and resources. Find...Catalytically inactive CRISPR plasmids for purification of target loci. CRISPR... CRISPR Purify CRISPR: Purify Genomic ... using CRISPR. Created with BioRender.com. Browse, sort, or search the tables below for CRISPR plasmids...
  5. Hot Plasmids - August 2020

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    ...find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources page...Addgene #140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using...-1 The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR plasmids. To find... Find the plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting... Find the plasmids at Addgene! CRISPR RNP electroporation and AAV donor infection (CRISPR-READI) enables...all-in-one dCas9-KRAB and dCas9-VPR targeting plasmids can be found at Addgene. Check out these plasmids for ...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...
  6. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    ...dystrophy. While mini-dystrophin expression levels were lower than expression of wild-type dystrophin in normal...sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains: 400bp homology arms to the...Drawbacks: Transgene expression varies with this approach, ranging from ~10% expression in primary human ...this AAV successfully mediated transduction and expression of the full-length transgene in vitro and in ...genomes complement each other to restore the full-expression cassette. AAV’s natural ability to reassemble...increasing the size of the AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split...overlapping split vector has equal or higher transgene expression when compared to using a single over-sized vector...
  7. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

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    ...Read all CRISPR blog posts Resources on Addgene.org Browse our CRISPR interference plasmids Find bacterial...bacterial CRISPR plasmids Read our CRISPR guide ...transposon system. This system has two parts: a plasmid expressing the Tn7 transposition genes (tnsA, tnsB, ...Mobile-CRISPRi into bacteria. Top: A Tn7 transposon containing the CRISPRi components and a plasmid with...different versions of dCas9. Following integration, Mobile-CRISPRi is stably expressed with no selection ...ready to move Mobile-CRISPRi into your bacteria of interest? Find Mobile-CRISPRi plasmids at Addgene! References...about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko, CRISPRi, and CRISPRa...
  8. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

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    ...of interest has been cloned into an expression plasmid, the plasmid can be introduced into host cells, ...using synthetic genes that are typically expressed from a plasmid or from an integrated sequence in a stable...that the plasmids and sequences for antibodies are not shared. For instance, through open plasmid sharing...sharing, the CRISPR community has created hundreds of useful variants of Cas9. By providing access to the ...Bradbury et al., 2018). Hybridomas can lose gene expression of the mAb-encoding genes or fail to revive after...Recombinant monoclonal antibodies are encoded in plasmids and produce a single antibody recognizing one ...heavy chain genes of the antibody are encoded in a plasmid. (Right) An assembled antibody protein binds an...
  9. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

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    ...lactose released activates expression of GFP from the lac operon. GFP expression can then be used to determine...fantastic tools it’s now easy to monitor protein expression, localization, and protein-protein interactions... Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs...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...
  10. Pooled Library Amplifications

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    ...recombined plasmid to overtake full-size plasmids (or for smaller plasmids to overtake larger plasmids, if the...Protocol (DOCX, 17 KB)   For CRISPR libraries, Genome-scale CRISPR-Cas9 knockout and transcriptional ...How many unique plasmids does your library contain?), the variance of size of plasmids in your library...Amplification Type Link to Protocol CRISPR Knockout Human CRISPR Knockout Pooled Library (Brunello) ...Amplification Protocol (PDF, 88.2 KB) CRISPR Knockout Human CRISPR Knockout Pooled Library (GeCKO v2) ... Protocol (DOCX, 15 KB) CRISPR Activation Human Genome-wide CRISPRa-v2 Libraries 104,540–209,080...Re-Amplification Protocol (PDF, 71 KB) CRISPR Activation Human Subpooled CRISPRa-v2 Libraries 12,285–20,145 ...
  11. AAV Molecular Tools

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    ...encode Cas9 nuclease used in the CRISPR-Cas9 system for targeted genome editing. ID Name Expression System...AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA CMV-driven, constitutive Expression of human codon-optimized Cas9 from Staphylococcus...with tetracycline (tet)-inducible expression systems . ID Name Expression System Activity Serotype PI 99118...transsynaptic tracer protein to express in presynaptic glutamatergic neurons that express Cre, Flp payload. Must...Cre-dependent expression of ER-localized TurboID (with HA and V5 tag). 8 Jonathan Long Overexpression Tools These...These AAV encode various genes for overexpression. ID Name Expression System Activity Serotype PI 58376...99118 pAAV-CAG-tTA CAG-driven, constitutive Expression of the tet-off transactivator (tTA). 2 Viviana Gradinaru...
  12. AAV Q&A with Tim Miles

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    ... of CRISPR/Cas9 Tips for a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids Adenovirus...for model organisms.    Q11: How consistent is expression is when injecting CAP-B10 or CAP-B22 in marmosets.... The continued development of more and smaller CRISPR tools is an exciting area for future AAV application...
  13. Tips for arabidopsis transformation

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    ...plant plasmid resources at Addgene  Choosing an expression system Choosing an appropriate expression vector...Addgene blog Engineering the plant genome using CRISPR/Cas9 Using pSiM24 for simplifying plant genetic engineering....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...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...for visual selection of transformants based on expression of RFP in the seed (Emami, Yee and Dinneny, 2013...
  14. Overcoming the Challenges of Lentiviral Production

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    ... multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually...encodes VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer...Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...and packaging plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection...high titer virus for transfer plasmids with small inserts (such as CRISPR guides) while a single 72h harvest... a usable titer, or fine-tuning selection and expression in your target cell line, researchers often find...By dividing the viral elements across multiple plasmids, the risk of recombination and consequently, the...
  15. Adenoviral Vector Production and Troubleshooting

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    ... primer for beginners Adenoviral delivery of CRISPR/Cas9 5 tips for troubleshooting viral transductions...death of transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of ...capacity is limited to 8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune response... capacity (up to 36 kb!), long-term transgene expression and negligible toxicity. However, their production...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...
  16. Luminescent Imaging with Nano-lanterns

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    ...for endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent signal for imaging...has a low quantum yield, and when transiently expressed in cells, generates much less signal than comparable...: 1262. PMID: 23232392 PMCID: PMC3535334 Find plasmids from this paper at Addgene. Takai, Akira, et ...4352-6. PMID: 25831507. PMCID: PMC4394297 Find plasmids from this paper at Addgene. Suzuki, Kazushi, ... 13718. PMID: 27966527 PMCID: PMC5171807 Find plasmids from this paper at Addgene. Additional Resources...All Our Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein ...
  17. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

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    ...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation...recombination, RFP is expressed. Cre can mediate one of two deletions to allow YFP or M-CFP expression; these deletions...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...ways to spatially and temporally control gene expression, especially in in vivo models, and new Cre-lox... of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need a quick primer on how...fluorescent protein in the construct, OFP (orange), is expressed when Cre is absent; stochastic Cre excision results...
  18. Distribution to Industry

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    ...Mutation Libraries CRISPR Thorsten Stiewe Use these donor vector libraries for CRISPR/Cas9-mediated saturation...collection of plasmids that are available to industry. Learn more about the available plasmids and how to...Distribution to Industry Featured Collections Search Plasmids Plasmids Kits Pooled Libraries Antibodies Ordering ...additional filters such as expression, vector type, species, and more. Plasmids Browse the table below to...see all of the plasmids that are currently available to industry scientists. ID Plasmid Gene/Insert PI...assembly. MoClo Toolkit Plant Expression Sylvestre Marillonnet 95 plasmids for cloning and assembling standardized...Fusions from Promega COVID-19 SARS-COV-2, ACE2, and CRISPR tools Fluorescent Proteins from the Michael Davidson...
  19. Quick Guide to All Things Lentivirus

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    ...genome and stable expression of the transgene   Transfer plasmids The transfer plasmid is the vector you...vectors fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral...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... facilitates nuclear export) is expressed from a separate plasmid, and the promoter of the 5’LTR has been...(tTA, also expressed by your transfer vector) binds to the TRE and activates the expression of your gene...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...
  20. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    ...requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene...enChIP Plasmids? All Hodaka Fujii lab plasmids Sign up to receive email alerts when new plasmids from ...vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system enables us to express the CRISPR... shopping for a wide range of plasmids. The merit of depositing plasmids to Addgene has been spectacular...IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita T, Fujii H. PLoS One. ... molecule technologies such as TAL proteins and CRISPR became available, I was very excited because genome-editing...recently developed a retroviral enChIP system using CRISPR technology. Can you describe this technology and...
Showing: 161 - 180 of 208 results