Skip to main content
Addgene
Open Menu

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

Information

In Category

We narrowed to 150 results for: cas9 expression vector

Showing: 121 - 140 of 150 results

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

    Type
    Blog Post
    ...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region...considerations, such as the starting material (DNA-based vector or purified RNA) and the initial state of the dsRNA... easiest approach to induce RNAi involves the expression of long hairpin RNA (hpRNA). This method is facilitated...construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene target sequence... Although downstream work is required, several vectors can be found at Addgene that could serve as an ...an excellent start (#22988, #137884). Both vectors allow the creation of amiRNAs based upon miRNA precursors...Bassik MC (2016) Systematic comparison of CRISPR/Cas9 and RNAi screens for essential genes. Nat Biotechnol...

  2. Lentivirus Production

    Type
    Protocol
    ...Lentivirus Production You may also like... Viral Vector Guides Mol Bio Protocols Introduction This protocol...be used to produce lentivirus from a lentiviral vector transfected into 293T cells using a polyethylenimine...Figure 1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI...per 10 cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would...

  3. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...compatible Y2H vector - BD pRPR1_gRNA_handle_RPR1t - gRNA expression empty vector pFA6a vectors - Epitope ...lentiviral vector for ORF expression pDest-565 - Gateway destination vector for bacterial expression; His,GST...gene expression vector URA3 Yeast pAG306GAL-ccdB - Yeast Gateway expression vector (see...Inducible Expression page Bacteria Lac, T7, araBAD, trp mTagBFP2-pBAD - Protein expression vector with C-...versions of pFastBac LIC vectors and pFastBac Dual LIC vectors for insect expression Fluorescent proteins ...Bacterial expression of EBFP2 blue fluorescent protein 35S-eGFP-nosT - Transient expression vector for eGFP...tag; 2NLS; pCDNA3 vector backbone, mammalian expression pENTR4-myc-nuc - pEntry vector that adds a C-terminal...

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

    Type
    Blog Post
    ...   It’s not an overstatement to say that CRISPR/Cas9 editing is one of the key tools now providing access...continues to be challenging. Organism specific viral vector systems would also be beneficial for many systems...snail can be manipulated by injected RNA and can express fluorescent proteins. This lab has created an automated...-Mediated Ovary Transduction of Cargo)  delivers Cas9 RNP to the arthropod germline by injection into ...

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

    Type
    Blog Post
    ...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...ways to spatially and temporally control gene expression, especially in in vivo models, and new Cre-lox...fluorescent protein in the construct, OFP (orange), is expressed when Cre is absent; stochastic Cre excision results... and inversion. The Brainbow-2.1 construct can express one of four colors (n-GFP, RFP, YFP or M-CFP.) ...remaining cassette can invert as long as Cre remains expressed; once Cre is removed from the system, the fluorescent...coding sequence is closest to the promoter will be expressed. This system differs from Brainbow-1.0 in that...

  6. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ... of expression. For example, CAG is a strong promoter that drives very high, ubiquitous expression, while...such as SaCas9, CRISPR gRNAs, or your gene of interest. Promoter The promoter drives the expression of the...outside of the ITRs does not get packaged in the AAV vector. The cloning capacity between the ITRs is ~4.4 ...type-specific, which can help you restrict the expression of your transgene to a specific species or location...to spatially and temporally control transgene expression. Lox sites are directional 34 bp sequences that...nuclear export of mRNA and helps increase the expression of your transgene. The WPRE is located at the...design a construct for the successful and specific expression of your gene of interest. References Haery ...

  7. CRISPR Plasmids - Parasites

    Type
    Collection
    ...gRNA plasmids, visit Addgene's Empty gRNA Expression Vectors page and search the table for "Parasite"....CRISPR expression plasmids that have been designed for use in parasites. CRISPR...research for decades. Recent developments with CRISPR-Cas9 gene editing have propelled both reverse and forward...forward genetics of these parasites. Moreover, CRISPR-dCas9 gene regulation has also been established for some...

  8. Hot Plasmids - August 2020

    Type
    Blog Post
    ...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...piggyBac vectors for uniform and sustained expression of single or multiplexed gRNAs and of dCas9 fusions...and track cells expressing both dCas9 and gRNAs. The multi-gRNA piggyBac vector integrates into ...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...#140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction... contain an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9-VPR repressor in hPSCs is also... the mCitrine gene. mCherry is constitutively expressed. Cells are then cultured and FACS sorted. Image...

  9. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...SELEX, the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex. Broccoli...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...

  10. Botman-Teusink Yeast FP Collection

    Type
    Collection
    ...Y., Nielsen, L. K. (2013). Dual gene expression cassette vectors with antibiotic selection markers for...Markerless Yeast Localization and Overexpression (MyLO) CRISPR-Cas9 Toolkit Other Addgene Plasmids Bas...Botman et al., 2019 . FP Tagging Yeast tagging vectors to create fusions of proteins of interest with ...yeast codon-optimized fluorescent proteins. These vectors are based on the pFA6a-link plasmid (Sheff et al...FPs in yeast. Vectors are based on the pDRF1 plasmid (Loqué et al., 2007) containing the PMA1 promoter and...blue, green, and red fluorescent protein tagging vectors for S. cerevisiae. PLoS One, 8 (7):e67902. https...Marker Tags Publication FP Overexpression Plasmids for constitutive overexpression of codon-optimized FPs ...

  11. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ... vitro, and can be used for measuring protein expression or localization in cells, or even single-particle...Mol Syst Biol. 2014 Galit Lahav EYFP Gateway vectors with EYFP split at 175 Arabidopsis homolog of the...Split-horseradish peroxidase (Alice Ting Lab 2016)  Split-Cas9 for inducible genome editing (Feng Zhang Lab 2015...

  12. Distribution to Industry

    Type
    Collection
    ...while also adding additional filters such as expression, vector type, species, and more. Plasmids Browse ...CRISPR Thorsten Stiewe Use these donor vector libraries for CRISPR/Cas9-mediated saturation genome editing...for Golden Gate assembly. MoClo Toolkit Plant Expression Sylvestre Marillonnet 95 plasmids for cloning...

  13. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...of interest when expressed at endogenous levels (Laflamme et al., 2019). CRISPR-Cas9 approaches are commonly...flow cytometry, and mRNA expression.   If your protein of interest is expressed at similar levels in ...useful to produce a sample with mixed expression, with the expression measurable through an epitope tag such...epitopes. Detection of overexpressed (tagged or untagged) protein When expression plasmids are available...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...caused by overexpression. Additionally, if the cell line used has endogenous levels of expression, this can...preferred. The overexpression approach is therefore best suited to cell lines without any expression of the target...

  14. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ... plasmids expressing Cre recombinase and filter the results by expression system and vector type. Find... GFP Plasmids 101: FLEx Vectors Optimizing AAV DIO and FLEx Vector Expression Plasmids 101: Knockout/Knock-In... plasmids expressing Flp recombinase and filter the results by expression system and vector type. Find... plasmids expressing Dre recombinase and filter the results by expression system and vector type. Find...genome or control gene expression or skip ahead to browse highlighted plasmids expressing Cre , Flp , Dre ,...Recognition Sites. Controlling Recombinase Expression By expressing the enzyme at specific times or locations...popular recombinase-dependent gene expression strategies. Gene Expression: A stop codon flanked with loxP...

  15. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...loxP sites such that their expression is turned on or off in cells expressing Cre. The result of a system...allows the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell...immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes...efforts have been made to create transgenic mice expressing fluorescent protein reporter constructs in the...above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after...other technologies such as optogenetics and CRISPR/Cas9 will help immunologists create even better tools...Subcellular Localization Find Fluorescent Protein Empty Vectors ...

  16. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...bacteria that have taken up this vector, but it won’t tell you whether the vector contains the gRNA insert or...X-gal allows you to identify cells that took up the vector and distinguish between plasmids that contain the...𝛽-galactosidase. Complementing this mutation by expressing this deleted portion, for example on a plasmid...functional 𝛽-galactosidase. Blue-white screens rely on vectors where the gene of interest is inserted into the..., you will see two bands: one representing your vector backbone and the other representing the insert....doesn’t contain the insert, you will just see the vector backbone band. Or, you can choose an enzyme that...specific to your insert or by using primers that are vector specific but amplify a potential insert. If you...

  17. Tetracycline Inducible Expression

    Type
    Collection
    ...popular empty backbones, expression vectors for transactivators, and vectors for other applications, or...Lentiviral Tet-Off vector for repressible expression with blasticidin selection. *Note: this vector contains an...PB-EF1a-TetOn3G PiggyBac vector expressing Tet-On 3G transactivator constitutively expressed by EF1a promoter ...Church 87360 TLCV2 Lentiviral vector for tet-inducible Cas9-2A-EGFP expression. Based on LentiCRISPR v2 ....Tetracycline (Tet) Inducible Expression Tetracycline (Tet) Inducible Expression Tetracycline Off Tetracycline...factors to further minimize background expression. Improved expression of the transactivator proteins in mammalian...Experimental Considerations Expression System If your gene of interest should only be expressed occasionally, use...

  18. Bacterial Expression Systems

    Type
    Collection
    ... Plasmid Collections Bacterial Expression Systems Bacterial Expression Systems Protein Purification Protein...bacterial expression systems. Read below and browse our curated collection of bacterial expression plasmids...Addgene Blog E. Coli Strains for Protein Expression Protein Expression Epitope Tags Protein Tags Return to ...to top Controlled Expression It isn’t always advantageous to constitutively express your gene of interest...turn on expression only at a specific time. This collection contains plasmids whose expression levels ...Addgene Controlled Expression Resources Check out our Tetracycline (Tet) Inducible Expression Collection for...systems. ID Plasmid Promoter Inducer Expression Species PI 44249 pdCas9-bacteria pTetO Anhydrotetracycline...

  19. Plan Your Experiment

    Type
    Guide
    .../or Cas9 expression or (2) a lack of efficient target cleavage in cells that do express both Cas9 and ...Figure 1). You will decide how to express Cas9, the delivery system for Cas9 and the guide RNA (gRNA), the...depositors ). Browse empty gRNA expression vectors . Select Your Expression and Delivery System Once you ...Delivery Using Viral Vectors Viral vectors are a great option, as they offer high expression levels, long-term...about viral vectors and their production, see our viral vector guides . Lentiviral vectors Lentiviral ...lentiviral preps . AAV vectors AAV vectors offer some advantages for viral vector delivery. Both lentiviral...non-homologous end joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity...

  20. CRISPR Guide

    Type
    Guide
    ...more about Cas9 multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included in ...a 2-vector system, you’ll need to either co-infect with a Cas9-producing lentivirus or use a Cas9-expressing...strand SpCas9-HF1 — disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 — increase Cas9 proofreading...more Cas9 fusion tools later in the guide. Figure 3: Overview of Cas9n. Increasing fidelity of Cas9 Researchers...enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined...
Showing: 121 - 140 of 150 results