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We narrowed to 111 results for: cas9 expression vector

Showing: 101 - 111 of 111 results

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

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...   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 ...

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

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ... 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 ...

  3. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...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...

  4. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...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...

  5. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...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...

  6. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ...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 ...

  8. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...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...

  9. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...much smaller proteins than SpyCas9, allowing them to be packaged in AAV vectors, potentially with multiplexed...al., 2024). In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study...showed that AAV-delivered CHARM could silence the expression of prion protein in mouse brains for at least...gene product; it simply turns the target protein expression off. Plus, DNA methylation is a long-lasting ...-silencing CHARM variants that stop their own expression after silencing their target — potentially improving...by targeting marker proteins that are uniquely expressed in a given cell type. For example, glial fibrillary...GFAP) is a glia-specific intermediate filament expressed in astrocytes throughout the brain, making it ...

  10. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...We generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP...plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene: Your ...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary mouse... IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita T, Fujii...

  11. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ... DNA compaction, an enhancer can increase the expression of a gene that is millions of base pairs away...Putting it all together, we get the following expression for step 1: (H3K4me1 + H3K27ac) – H3K4me3 The...enhancers with CRISPR Armed with the genome math expression for NANOG enhancers, we’re ready for step 2: ...of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus in the same family as...regions and their reverse complements were parsed for spCas9 PAM sites (NGG) and then filtered based on two ...
Showing: 101 - 111 of 111 results