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We narrowed to 224 results for: cas9 expression plasmid

Showing: 161 - 180 of 224 results

  1. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ... Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors ... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western blotting...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...isolate individual cell clones with different expression characteristics from a population of transfected...Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org...

  2. CRISPR Plasmids - Zebrafish

    Type
    Collection
    ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...need Cas9 plasmid Chen and Wente 42250 DR274 T7 BsaI In vitro transcription none, need Cas9 plasmid Joung...than NHEJ. ID Plasmid Gene/Insert Vector Type Promoter PI Publication Empty gRNA Expression Vectors Select...Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When using...nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with a ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...CRISPR plasmids that have been designed for use in zebrafish. CRISPR...

  3. CRISPR Plasmids - Yeast

    Type
    Collection
    ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert...cerevisiae LEU2 none, need Cas9 plasmid Lu Do you have suggestions for other plasmids that should be added ...of the coding sequence. If the plasmid you’re using does not also express a gRNA, you will need to use ...sites. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Empty gRNA Expression Vectors Select... Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When ...

  4. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...selectable marker, and whether the plasmid contains Cas9. ID Plasmid Expression System Promoter Guide RNA Type...below for CRISPR prime editing plasmids. Plasmids are available for expression in mammalian systems, bacteria...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Drosophila ID Plasmid Gene/Insert...Type Cloning Enzyme Co-expressed Cas9 Selection PI 132777 pU6-pegRNA-GG-acceptor Mammalian hU6 pegRNA ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...prime editor plasmids for (search and replace) genome editing. CRISPR...

  5. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...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... editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “... plasmid... system that uses modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety of customizable...

  6. CRISPR Plasmids - Base Edit

    Type
    Collection
    ...below for CRISPR base editing plasmids. Plasmids are available for expression in mammalian systems, bacteria...are created by fusing Cas9 nickase or catalytically inactive “dead” Cas9 (dCas9) to a cytidine deaminase...CRISPR Plasmids: Base Edit Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit...editors are available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have ...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast ID Plasmid Gene/Insert...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...

  7. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...Addgene plasmid ID 42230). This plasmid allows for the simultaneous expression of sgRNA and SpCas9, but ...cloning and expression purposes. NOTE: Here we discuss the usage of the pSpCas9(BB) plasmid (pX330) (Addgene...utilized, such as pX458 (Addgene plasmid ID 48138) or pX459 (Addgene plasmid ID 48139), which include GFP ...constructs in which multiple sgRNAs may be expressed from a single plasmid. Add “CACC” before the 20-mer guide...This protocol involves the delivery of CRISPR/Cas9 plasmids by electroporation 4 . This protocol is described.... However, transient transfection of CRISPR/Cas9 plasmids may be successfully adapted to numerous cell...Add 5 μg of each CRISPR/Cas9 construct (10 μg total). Add 0.5 μg of GFP expression construct. Electroporate...

  8. CRISPR Plasmids - Mammalian Expression

    Type
    Collection
    ...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert...CRISPR Plasmids: Mammalian Expression The following CRISPR plasmids have been designed for use in mammalian...FLAG-tagged Cas9. In the CAPTURE system, Cas9 is tagged with a biotin acceptor site and co-expressed with BirA...NHEJ. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Browse CRISPR Plasmids By Function...repair (HDR). ID Plasmid Gene/Insert Promoter Selectable Marker Publication Prime Edit Cas9 H840A nickase...of the coding sequence. If the plasmid you’re using does not also express a gRNA, you will need to use ...

  9. CRISPR Plasmids - Tagging

    Type
    Collection
    ...Tagging Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA...and Myers Tagging Plasmids Protein Species Tag Donor Plasmid gRNA plasmid gRNA plasmid FLAG pFETCh_Donor...the remaining 5 plasmids include Cre and FLP expression vectors, a general cloning plasmid, and a prebuilt...Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_T2 (Addgene #79888) , which expresses an untagged Cas9 AND a gRNA... This CRISPR/Cas plasmid-system consists of two components: A vector containing Cas9 and a validated gRNA.... Multiple gRNA plasmids may be used. The HDR (homology directed repair) donor plasmid with homology arms...well as pX330S-2-PITCh, expressing a generic gRNA targeting the PITCh donor plasmid. Note that the principle...

  10. Brzezinski Lab CRISPR Collection

    Type
    Collection
    ...and 175574 express dCas9-KRAB-MeCP2 for repression. Plasmids 175571 and 175572 express dCas9-VPR for activation...shuttle plasmid for shuttling U6-guide cassettes to make dual guide expressing plasmids dCas9-KRAB-MeCP2...guides can be expressed from the same plasmid by utilizing one of the dual guide shuttle plasmids to insert...enhancer sequence repression dCas9-VPR fusion for gene activation CRISPR/Cas9 Plasmid Variants Figure 1: Schematic...advantage of the Zhang lab all-in-one (guide/Cas9/reporter) pX458 plasmid family. They have made several modifications... the CRISPR Collection. Plasmids 159654 and 159655 are single guide expressing double stranded break constructs...constructs. Plasmids 171098 and 175570 allow conversion to a dual guide construct. Plasmids 175573 and ...

  11. 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...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...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 ...

  12. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...has a low quantum yield, and when transiently expressed in cells, generates much less signal than comparable... endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent signal for imaging ...: 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 ...

  13. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ...interest (or the whole genome), they knock down gene expression temporarily. RNAi libraries are especially useful... Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166): 80-84 (2014). doi: https...

  14. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...Browse fluorescent protein plasmids and resources See all Jaffrey lab plasmids ... cellular processes by transforming cells with plasmids encoding aptamer-tagged RNA and later introducing...aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment...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...SELEX, the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...

  15. CRISPR Plasmids - Plants

    Type
    Collection
    ...of Cas-containing plasmids. ID Plasmid Promoter Cloning Enzyme Co-expressed Cas9 Cas9 System Selection ...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert...PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such as ...CRISPR expression plasmids that have been designed for use in plants. CRISPR...repair (HDR). ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Prime Edit Cas9 H840A nickase...of the coding sequence. If the plasmid you’re using does not also express a gRNA, you will need to use ...

  16. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...generation of multiple edits or implementation of expression systems to accurately mimic a disease phenotype...CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of the genome Tips for a first...resources on Addgene.org CRISPR Guide CRISPR Plasmids and Resources CRISPR History and Development ...

  17. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...High fidelity Cas9s: Three high fidelity Cas9 variants , SpCas9-HF, eSpCas9, and HypaCas9 display very ...III and Cas9 enzymes, which together separate the individual crRNAs. (3) The tracrRNA and Cas9 nuclease...posts , or find CRISPR plasmids for your research. Addgene has CRISPR plasmids for the following models...Staphylococcus aureus (SaCas9) is smaller than SpCas9 and more easily packaged in AAV. Cas9 orthologs also have...beyond SpCas9, moving us closer to being able to target every locus in every genome. Multiple Cas9 orthologs...capability of Cas9 to make other types of modifications at specific loci. Activate or Repress : dCas9 fused to...known as CRISPR arrays. Separate Cas proteins then express and process the CRISPR loci to generate CRISPR ...

  18. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...Turquoise (CeNL; Plasmid #85199), Yellow (YeNL; Plasmid #85201), Orange (OeNL; plasmid #85202) and Red ... Researchers have been sharing plasmids ever since there were plasmids to share. Back when I was in the...specific strategies and tactics for getting the plasmids he wanted. Addgene makes it so much easier to ...endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc is very inviting for endogenous...kDa. An example is the CRISPaint-NLuc construct (Plasmid #67178) for use in the system outlined in Schmid-Burgk...reporter was used to sort successfully infected/expressing cells by flow cytometry. The isolated cells were...CyOFP1-NLuc-CyOFP1; Antares). Direct injection of plasmids for Antares or firefly luciferase into mouse livers...

  19. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...dystrophy. While mini-dystrophin expression levels were lower than expression of wild-type dystrophin in normal...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...overlapping split vector has equal or higher transgene expression when compared to using a single over-sized vector...could have a big impact on the rate of transgene expression with an overlapping AAV since both of these variables...sequences, the group achieved ~80% of muscle cells expressing mini-dystrophin.  3. Hybrid  Overview: The hybrid...

  20. CRISPR Plasmids - dCas9-FokI

    Type
    Collection
    ...tables below for CRISPR-FokI plasmids. Plasmids are available for expression in mammalian systems and Drosophila...Drosophila. Mammalian ID Plasmid Gene/Insert Promoter PI Publication Drosophila ID Plasmid Gene/Insert Promoter...Yeast ID Plasmid Gene/Insert Promoter PI Publication Do you have suggestions for other plasmids that should...CRISPR plasmids with dCas9 fused to FokI for creating double-strand breaks (DSBs). CRISPR... CRISPR: dCas9-FokI Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit Transcriptional...Tools CRISPR Blog Posts A catalytically inactive Cas9 (dCas9) is fused to FokI nuclease. When FokI dimerizes... be added to this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve...
Showing: 161 - 180 of 224 results