Skip to main content
Addgene

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

In Category

We narrowed to 716 results for: CaS;

Showing: 161 - 180 of 716 results

  2. Modular Cloning Guide

    Type
    Guide
    ...efficiency intron-optimized SpCas9-coding gene. MoClo CRISPR/Cas Toolkit for Plants Genome Engineering, CRISPR...Expression Vladimir Nekrasov 95 plasmids including CRISPR/Cas nucleases, base editors, gRNA backbones, and promoters...CRISPR/Cas9 vectors expressing multiple gRNAs (up to seven) with a Cas9 nuclease, nickase, dCas9, or dCas9...mechanisms. zCas9i cloning kit: MoClo-compatible CRISPR/Cas9 cloning kit with intronized Cas9 Genome Engineering...assembly of CRISPR/Cas9 constructs for use in plants. The constructs are made with zCas9i, a high efficiency...create plant expression vectors containing several cassettes and generate multi-construct transgenic plants...biology and multigene engineering, including CRISPR/Cas9 components. MoChlo: Modular Cloning Chloroplast ...

  6. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ...     References and Resources References Ascoli CA, Aggeler B (2018) Overlooked benefits of using polyclonal...their ability to bind the target antigen. In this case, the serum is run over a column containing the target...

  7. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ... gene networks with an integrated RNA and CRISPR/Cas toolkit in human cells. Nissim L, Perli SD, Fridkin...In the short time since its development, CRISPR/Cas9 genome editing has been used to study the effect...recombination. To further increase the utility of CRISPR/Cas9, it will be necessary to improve its multiplexing...RNAs (gRNAs) are commonly packaged in 400-500 bp cassettes containing the RNA pol III promoter, gRNA and ...and pol III terminator. These relatively large cassettes (considering the gRNA itself is ~100 bases) limit...construct size, it requires coexpression of Csy4 with Cas9 and the polycistronic transcript. As nucleases may... (15% and 9% of target sites.) How many loci can Cas9 modify? To test the application of this system to...

  8. Chemogenetics Guide

    Type
    Guide
    ...hM1Dq hM5Dq Human M3 muscarinic G αq CNO* Increase Ca 2+ Neuronal burst firing Armbruster et al., 2007 ...1670767 (Link opens in a new window) Tung JK, Gutekunst CA, Gross RE. (2015). Inhibitory luminopsins: genetically-encoded...

  9. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell,163(3), 759-771. PubMed PMID: 26422227...Comparing the Cas9 and Cpf1 CRISPR Nucleases   Cas9 Cpf1 Components crRNAtracrRNACas9 crRNACpf1 ... guide page for a review of Cas9.  For a brief comparison of Cpf1 vs. Cas9, see the table below. Table... Endonuclease size spCas9: ~4 kbsaCas9: ~3 kb ~3.9kb crRNA/gRNA length gRNA: ~100 nt crNA: ~42...promoters).  spCas9 and its gRNAs are also larger than their Cpf1 counterparts. Table 2. Cas9 Multiplexing...Lentiviral Expression Cassettes lentiviral transduction One vector expresses Cas9, eGFP, and up to 4 ... Requires co-transfection of Cas9 Nissim et alTsai et al PTG Cassettes transfection gRNAs expressed ...

  10. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...Loci in Living Human Cells by an Optimized CRISPR/Cas System.” Cell 155 (2013): 1379-1491. Pubmed PMID:... a colorful Cas9 toolbox, Ma et al. turned to SpCas9 and its orthologs NmCas9 and St1Cas9. Each ortholog...locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot induce gene editing, but it retains its...bright-field image. Image from Ma et al, 2016. Using dCas9 to visualize genomic loci in vivo The flexibility...By using a gRNA to target fluorescently labeled dCas9 to a given genomic locus, you can track the localization...to noise ratio of the system, they modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly...amount of background fluorescence from unbound GFP-dCas9. With these improvements, labeling efficiency was...

  11. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...References and resources References Bates A, Power CA (2019) David vs. Goliath: The Structure, Function...anti-IgG Fab fragments make good secondaries for cases where you need to pre-form primary-secondary antibody...antibody recognizes endogenous antigens. In these cases, even though one full antibody may be able to get...

  13. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ...libraries depend on many gRNAs delivered with a relevant Cas nuclease. MoClo can be used to assemble multiplexed... part, will behave in a cell. For unpredictable cases, MoClo provides two answers: first, sets of parts...

  14. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...They're used to determine the PAM requirements of Cas enzymes. CRISPR-ARE is a new system from Jiazhang...and editing with a single Cas9-VPR in Saccharomyces cerevisiae. To control Cas9 activity and influence DNA... editor targets DNA (via catalytically inactive dCas9) to modify chromatin. A fluorescent protein can ...CRISPR system developed by Taekjip Ha’s lab where Cas9-gRNA complex binds the target DNA without cleavage...cleavage until it's activated by light. Once activated, Cas9 cleavage can induce DSBs within minutes. New CRISPR...Jacobs-Wagner’s lab produce different levels of dCas9 expression and contain different antibiotic resistance... lab developed a chemogenetic system that brings Cas9 to ubiquitin ligase for degradation.      New...

  15. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...Central PMCID: PMC2593125. 4. Saunders A, Johnson CA, Sabatini BL. 2012. Novel recombinant adeno-associated... switch conditional knock-in cassette. As shown in Figure 2, the cassette should contain a reporter DNA...For this FLEx switch to successfully work, the cassette would need to contain the BFP coding sequence ...antisense orientation (Figure 1, top). The entire DNA cassette would be flanked by two pairs of target sites-...in the opposing orientation, you have created a cassette that will express GeneR only in the presence of...recombination event would excise the reporter from the cassette, allowing for expression of GeneR. This allows...

  16. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...contributed by Derek Jacoby from Makerspace Victoria, CA Over the decades science has become increasingly ...with  Addgene. We now have the incredible CRISPR/cas9 tools at our disposal to to modify the worm genome...be useful to our model  so that we can use CRISPR/cas9 to build the mutants, analyse their motor behaviour...project, but the primary goal is learning in this case.   Funding Makerspaces One of the main challenges...- everyone contributes each month - with only occasional supplementation with grant funds, money is always...

  17. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...fast, and cheap distribution of the newest CRISPR-Cas plasmids, has been a key driver of the technology...complete without the inclusion of some interesting case studies, and Zimmer provides several. In Part Three...winners for developing the technology. Though these case studies are “old,” they lead Zimmer to one of his...research, how they conduct their research, and in some cases, if they conduct their research. Even if Zimmer ...

  19. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...pieces of RNA that bind to a cell’s DNA, telling the Cas machinery where to cut.      Fig. 2: Heatmap... the guide from binding to the genome or prevent Cas9 from cutting, both of which can introduce significant... are undercounted by approximately 20%. In some cases, this may lead researchers to deprioritize genes...

  20. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...anti-CRISPRs were identified that inhibit the S. aureus Cas9, a Cas protein that is smaller and easier to deliver...CRISPR/Cas9 constructs. zCas9i is a high efficiency intron-optimized SpCas9-coding gene. SlugCas9 is a ...promoters driving expression of Cas9 were used in this study.  The zCas9i cloning kit is a plant cloning... of the population.  Because indel generation by Cas9 can occur at a steady rate, scientists found a way...a compact Cas9 ortholog from Staphylococcus lugdunensis that recognizes a NNGG PAM and has comparable ...comparable activity to SaCas9. A new yeast inducible CRISPRi library - Ampl133 targets S. cerevisiae genes with...
Showing: 161 - 180 of 716 results