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We narrowed to 165 results for: cas9 expression vector

Showing: 121 - 140 of 165 results

  1. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...contains Cas9. ID Plasmid Expression System Promoter Guide RNA Type Cloning Enzyme Co-expressed Cas9 Selection...Publication Empty Prime Editing gRNA Vectors A selection of empty gRNA vectors suitable for prime editing are...below. Use the search bar to find a gRNA vector based on expression system, promoter, the type of gRNA (e.g...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian...transcriptase (RT) is fused to the C terminus of Cas9 H840A nickase. The fusion enzyme is capable of installing...template. During prime editing, the pegRNA directs the Cas9 nickase to the target sequence where it nicks the...nuclear localization signals, and two mutations in Cas9 to enhance nuclease activity; can be used in PE2...

  2. CRISPR Plasmids - Tagging

    Type
    Collection
    ...Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA, & FBL-specific...targeting vector assembly, and the remaining 5 plasmids include Cre and FLP expression vectors, a general... one vector from the Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_T2 (Addgene #79888) , which expresses an untagged...plasmid-system consists of two components: A vector containing Cas9 and a validated gRNA, based on the Zhang...consists of two components: An all-in-one CRISPR/Cas9 vector based on the Zhang lab's PX330 , simultaneously...gene-specific CRISPR and donor vectors as well as pX330S-2-PITCh, expressing a generic gRNA targeting the... that uses PCR to generate both an expression cassette for the Cas9-programming sgRNA and HR donors for...

  3. 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 ...

  4. CRISPR Plasmids - Yeast

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    Collection
    ...Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such...Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG)...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-repressor...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian... Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA...immunoprecipitate FLAG-tagged Cas9. Design your gRNA sequence to direct dCas9 to a specific locus, avoiding...

  5. CRISPR Plasmids - Mammalian Expression

    Type
    Collection
    ...FLAG-tagged Cas9. In the CAPTURE system, Cas9 is tagged with a biotin acceptor site and co-expressed with BirA...Publication Empty gRNA Expression Vectors You can use the tables on Addgene's Empty gRNA Vectors page to search...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-repressor... CRISPR Mammalian Expression CRISPR Plasmids: Mammalian Expression The following CRISPR plasmids...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian... Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA...

  6. CRISPR Guide

    Type
    Collection
    ...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...new tools. Figure 3: Overview of Cas9n Nickase and dead Cas9 The two Cas9 nuclease domains, RuvC and HNH...enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined...

  7. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...the 20-mer is not G. sgRNA expression from the U6 promoter of the pX330 vector is enhanced by the inclusion...Add 5 μg of each CRISPR/Cas9 construct (10 μg total). Add 0.5 μg of GFP expression construct. Electroporate... for cloning and expression purposes. NOTE: Here we discuss the usage of the pSpCas9(BB) plasmid (pX330...plasmid allows for the simultaneous expression of sgRNA and SpCas9, but does not contain markers for selection...genomic deletions in mammalian cell lines using CRISPR/Cas9 CRISPR...and Daniel Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell lines...Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9. Bauer DE, Canver MC, Orkin SH. J. Vis. Exp. (95...

  8. AAV Molecular Tools

    Type
    Collection
    ...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... Viral Vector Packaging Service AAV Molecular Tools Viral Vector Packaging Service: Molecular...with tetracycline (tet)-inducible expression systems . ID Name Expression System Activity Serotype PI 99118...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...

  9. 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...

  10. 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...

  11. CRISPR Plasmids - Base Edit

    Type
    Collection
    ...are created by fusing Cas9 nickase or catalytically inactive “dead” Cas9 (dCas9) to a cytidine deaminase...editors are available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have ...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian...base editors, the evolved TadA domain is fused to a Cas9 protein to create the adenine base editor. Both ...have been made by optimizing expression of the fusions, modifying the linker region between Cas variant and... lab and CRISPR-X from the Bassik lab , in which Cas9 is fused to activation-induced cytidine deaminase... editing plasmids. Plasmids are available for expression in mammalian systems, bacteria, plants, yeast...

  12. CRISPR Plasmids - Plants

    Type
    Collection
    ...Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such...plasmids. ID Plasmid Promoter Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI 46968 pICSL01009::AtU6p...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-repressor...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian...CRISPR expression plasmids that have been designed for use in plants. CRISPR...Promoter Selectable Marker PI Publication Prime Edit Cas9 H840A nickase fused to a reverse transcriptase (...

  13. CRISPR Plasmids - dCas9-FokI

    Type
    Collection
    ...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian...Tools CRISPR Blog Posts A catalytically inactive Cas9 (dCas9) is fused to FokI nuclease. When FokI dimerizes...CRISPR-FokI plasmids. Plasmids are available for expression in mammalian systems and Drosophila. Mammalian... CRISPR dCas9-FokI CRISPR: dCas9-FokI...with dCas9 fused to FokI for creating double-strand breaks (DSBs). CRISPR...gRNAs, binding ~15-25 bp apart, are required for dCas9-FokI to dimerize at a given region of the genome...

  14. 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...

  15. 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...

  16. 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 ...

  17. CRISPR Plasmids - Cascade-Cas3

    Type
    Collection
    ...Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian...genomic deletions. Compared to deletions generated by Cas9, large deletions created using Cas3 have fewer off-target...underutilized compared to their Class 2 counterparts like Cas9. This is attributed in part to the difficulties ...getting all components of the Cascade complex to express equally. To combat this, some CRISPR-Cas3 plasmids...Cascade-Cas3 plasmids. Plasmids are available for expression in mammalian systems, bacteria, and plants. Mammalian...

  18. CRISPR Plasmids - gRNAs

    Type
    Collection
    ...gRNA expression vectors available from Addgene....Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian...species or PAM binding variant of Cas9. For instance, wild-type SpCas9 must be used with targets that are...increases off-target effect. Which species or variant of Cas9 ( S. pyogenes , S. aureus etc.) was this gRNA sequence...experimental use of these plasmids. ID Plasmid Gene/Insert Vector Type PI Publication Do you have suggestions for...

  19. 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...

  20. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
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    ...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...
Showing: 121 - 140 of 165 results