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We narrowed to 1,034 results for: LAS

Showing: 281 - 300 of 1034 results

  1. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...transfection of single plasmids vs pooled plasmids; for multiple edits, single plasmids are generally more...transfected on a single plasmid. This is versus 2.4% of cells transfected with a pool of plasmids that contained...sites. This eliminates bulk from crRNA expression plasmids resulting in smaller, more streamlined vectors...contained one crRNA per plasmid plus a Cpf1 expression vector.  See the graph in figure 2 for a comparison of...of a crRNA array that can be processed by Cpf1. Lastly, an AAV vector was used to multiplex edit 3-4 genes...your research? Check out the Cpf1 multiplexing plasmids available from Addgene. If you have any questions.... Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell,163(3), 759-771. PubMed...

  2. Kwabena Duedu on Public Health, Biofuels, and Doing Science in Ghana

    Type
    Blog Post
    ... awarded plasmids from Addgene as a successful applicant to the Addgene, Seeding Labs Plasmid Grant. For...Learn about the Addgene, Seeding Labs Plasmid Grant Read about Plasmid Grant Recipient, Gladys Kaba Resources... fellow nonprofit, Seeding Labs, to distribute plasmids to researchers in developing regions and thereby...accelerate their research. Kwabena came across the plasmid grant in Seeding Lab’s Newsletter and recently ...plans to use his research experiences as well as plasmids from Addgene to bring new opportunities to Ghanaian... Kwabena applied for the Addgene, Seeding Labs plasmid grant. 7:12 - 7:11 : Kwabena's work in Christopher... recent recipient of the Addgene, Seeding Labs Plasmid grant and is working on a variety of projects in...

  3. 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 ...kDa. An example is the CRISPaint-NLuc construct (Plasmid #67178) for use in the system outlined in Schmid-Burgk...CyOFP1-NLuc-CyOFP1; Antares). Direct injection of plasmids for Antares or firefly luciferase into mouse livers...Red (ReNL; plasmid #85203) NLuc fusions. The Nagai lab has additionally deposited more than 25 fusions of.... This construct is available through Addgene (Plasmid #83926). Live-Cell Voltage Sensor  Ingaki and ...

  4. What's New in CRISPR - March 2020

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    Blog Post
    ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...is inaccessible and the complex remains in the cytoplasm. Read the paper in Cellular and Molecular Bioengineering...Bioengineering Find the optogenetic repressor plasmids Cytosine base editors with minimized off-target...paper in Nature Biotechnology Find the base editor plasmids CRISPRi in Caulobacter crescentus Figure...once. Read the paper in mBio Find the CRISPRi plasmids A multiplexed gRNA piggyBac transposon system... Read the paper in Scientific Reports Find the plasmids Enhanced CRISPR-based DNA demethylation ...

  5. What's New in CRISPR - September 2019

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    Blog Post
    ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools... drug inducible Cas9 plasmids E. coli genome-wide inhibition library In the last edition of “What’s New...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...vector sequence. Homology arms that flank the donor plasmid cargo are complementary to either side of the double-stranded...domain needed for DNA unwinding. They’ve deposited plasmids for active and inactive CasX for E. coli and human...expression. Read the paper in Nature Find the CasX plasmids Drug inducible transcriptional activation and...estrogen receptor. This protein fusion stays in the cytoplasm, but with the addition of 4-hydroxytamoxifen, ...

  6. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... involving two-plasmid or single-plasmid systems (Vo et al., 2020). In the two-plasmid system, an effector...group’s initial report of VchINT, three separate plasmids and multiple promoters were used to express the...effector plasmid (pEffector) expresses all seven required genes and the CRISPR RNA under the same promoter...promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also be combined...combined in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and ...and the ability to easily cure cells of the plasmid after integration. Figure 2: (A) Streamlined ...insertions with Type I-F INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow...

  7. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...Originally published Nov 12, 2015 and last updated Aug 20, 2020. Cas9 can be used to modify any desired...284 . https://doi.org/10.1038/nature24049 Find plasmids from this paper at Addgene. Amrani N, Gao XD,... https://doi.org/10.1126/science.aaq0180 Find plasmids from this paper at Addgene. Kim D, Kim J, Hur...863–868 . https://doi.org/10.1038/nbt.3609 Find plasmids from this publication at Addgene. Kim E, Koo ...8: . https://doi.org/10.1038/ncomms14500 Find plasmids from this publication at Addgene. Kleinstiver...485 . https://doi.org/10.1038/nature14592 Find plasmids from this publication at Addgene. Kleinstiver...869–874 . https://doi.org/10.1038/nbt.3620 Find plasmids from this publication at Addgene. Hu JH, Miller...

  8. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ... scarless plasmid inserts.  Want to Learn More about DNA Assembly? Check Out Our Plasmids 101 eBook!...as easy as obtaining a plasmid with the gene of interest (search Addgene plasmids by gene here), designing...fragments (containing regions of homology) cut from plasmids via restriction enzyme digestion, and used directly...as simple as combining your favorite linearized plasmid with a synthetic oligo that contains a region of... desired edits to the target region. Assembled plasmids can be directly transformed into E.coli for plating...Other Long-Homology Based Cloning Methods Screen Plasmids by Colony PCR Addtional Resources on Addgene....Find Validated gRNAs Browse All Plasmids ...

  9. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...integrase-deficient packaging plasmid instead of the standard packaging plasmid. For more details on how to...advantage for researchers looking to generate long-lasting changes in cells. However, for some applications...lentiviral genome is distributed across multiple plasmids. For simplicity, we’ll use the second-generation...system as an example.  The lentiviral transfer plasmid encodes your insert of interest, flanked by long...this portion into the host genome. The packaging plasmid contains Gag, Pol, Rev, and Tat genes. Specifically...production of lentiviral particles.  The envelope plasmid typically encodes the vesicular stomatitis virus...into the host cell.  Transfecting these three plasmids into cells enables the production of lentiviral...

  10. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...e.g., sister chromatid, plasmid, etc.), and consists of three pathways: classical double-strand break repair...covered in detail here. Classical double-strand break repair (DSBR) In the classical DSBR pathway, the 3’...disrupted in the donor. Choosing a type of donor: plasmid, oligo, double or single stranded? The size of ...longer than 200 bases can be difficult, thus dsDNA plasmids are generally preferred for large insertions such...fluorescent proteins or selection cassettes. This class of template should have homology arms between 500...500-1000 bps. HDR efficiency with circular plasmid templates is generally low; to increase the frequency...edits, researchers have designed self-cleaving plasmids that liberate the targeting region from the vector...

  11. Antibodies 101: Isotypes

    Type
    Blog Post
    ...There are two IgA subclasses: IgA1 and IgA2.  You can learn more about isotype classes in our Antibodies...There are a lot of ways to classify antibodies: monoclonal, polyclonal, scFvs, nanobodies, and the list...switch isotypes? What is an isotype? An isotype is a class of antibody that’s determined by its heavy-chain...and α. Some isotypes have different versions or subclasses that have minor differences in their heavy chain...the hinge region in IgM antibodies. IgM has no subclasses and is found mostly in blood. IgD While IgD is...monomer structure with a valency of two and has no subclasses. IgG IgG is the most abundant isotype in blood...monomer with a valency of two. There are four IgG subclasses, numbered based on their abundance in blood: ...

  12. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...Interested in using PTG plasmids in your research? Check out the PTG plasmids or plant expression systems...indel generation for two PTG constructs in rice protoplasts. Compared to matching traditional gRNA-expressing...3 and 31 times higher, respectively. In rice protoplasts, Xie et. al observed slightly higher mutation...mutation rates than they’d previously reported in protoplasts (15% and 9% of target sites.) How many loci can...gRNAs targeting 4 MAPK genes, again using rice protoplasts. Each pair of gRNAs targeted genomic sites separated...common to prokaryotes and eukaryotes. Though the plasmids made by Xie et. al are specifically designed for....1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication at Addgene Other Techniques...

  13. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    ...stages and contains multiple cell types: fibroblasts, myofibroblasts, neuroendocrine, adipose, immune, inflammatory...cells are Cancer-Associated Fibroblasts, or CAFs. CAFs are altered fibroblasts and promote the development...gland biology and neoplasia 10.3 (2005): 249-260. PubMed PMID: 16807804. Hanahan, Douglas, and Lisa M. Coussens...Addgene.org Visit our cancer research plasmids and resources page  Find plasmids for your research Learn about...began focusing on the extracellular matrix and fibroblasts. With each cell mechanism discovered in microenvironments...Saidou, and Kounosuke Watabe. "Cancer associated fibroblasts (CAFs) in tumor microenvironment." Frontiers ...

  14. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET ...BirA* fusion proteins are expressed in cells via plasmids or by the creation of stable cell lines and these...interaction with the BirA* fused bait proteins ceases. Lastly, Split-BioID and BioID identify proteins that interact...Interested in using Split-BioID with your research? Plasmids are available from Addgene! References Schopp... Central PMCID: 5467174. Find the Split-BioID Plasmids Here! Munter, S.D., Görnemann, J., Derua, R.,...site than in Schopp et al. Find these Split-BioID Plasmids Here!   Burke, B., Kim, D.I., Roux, K.J., & Raida...PubMed Central PMCID: 3308701. Find the BioID Plasmids Here! Macneill, S., & Varnaitė, R. (2016). Meet...

  15. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...both NeuroMab and the Allen Brain Atlas. The comparison with two atlases enables us to see the reproducibility...publicly available 2D/3D mouse atlas from antibodies that we screen. This atlas will enable scientists to ...and uploading verified antibodies onto the mAb3D atlas website. This website can then be used by thousands...distinct imaged pattern should align with what the atlases (mentioned above) and literature suggest, providing...distinct imaged pattern does NOT align with the atlases/literature and is unrelated to the target protein...of Giving As an open-source database, the mAb3D Atlas will foster collaboration, progress, and innovation...closer to resolving the antibody crisis. The mAb3D Atlas is a public service to the science community, and...

  16. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...Cascade-Cas3 plasmids in Addgene's collection! A “complex” system Cas3 is part of the Class 1 CRISPR family...complexity. Figure 1: Schematic of Class 1 CRISPR types. Class 1 includes type I with a Cas3 effector...out the brunt of the mechanical work for editing. Class 1 CRISPR systems make up the majority of CRISPR-...CRISPR-based immune systems in bacteria and archaea. Class 1 is further broken down into different types, with...making up type I, the most abundant type in the Class 1 category. These type Is can then be further broken... Various labs have designed all-in-one Cascade plasmids — such as the Tomoji Mashimo lab and the Tilmann...Koonin, E. V. (2015). An updated evolutionary classification of CRISPR–Cas systems. Nature Reviews Microbiology...

  17. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...packaging plasmids: the pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing...with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly called...blog Parts of the AAV Packaging PlasmidParts of the AAV Transfer PlasmidAAV Variables That Matter AAV protocols...called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps of AAV production, iodixanol...repeats (ITR). You should always have a high-quality plasmid standard and rAAV reference material of a known...before undergoing titering, to remove residual plasmid DNA carried over from the production process. For...

  18. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...Fusions Plasmids 101: Secondary Nanobody Toolbox Resources on Addgene.org View the Antibody plasmid collection...The Seeger lab found a way around this by using plasmids to generate sybodies, synthetic nanobodies against...nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target epitopes that...form of mRNA for academic research. The template plasmids for generating your own libraries are also available...available in the Sybody Generation Toolbox plasmid kit: SB_concave, SB_loop, SB_convex. These scaffolds...efficiently subcloned into the phagemid pDX_init plasmid, using the fragment exchange (FX) cloning technique... subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins for further...

  19. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...Browse our CRISPR interference plasmids Find bacterial CRISPR plasmids Read our CRISPR guide ...transposon system. This system has two parts: a plasmid expressing the Tn7 transposition genes (tnsA, tnsB... tnsB, tnsC, tnsD, and tnsE), and a plasmid that contains the Mobile CRISPRi cassette flanked by left ...copy of the RP4 transfer machinery mobilize these plasmids for transfer to recipient cells during a tri-parental...Mobile-CRISPRi ICE element forms a conjugative plasmid-like DNA circle and is transferred to recipient...transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes are transferred...RP4 transfer machinery, which mobilizes the Tn7 plasmids, and lack the ability to synthesize diaminopimelic...

  20. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...lentiCas9-blast. A549 cells were transduced (MOI = 37) and selected with 1 µg/mL blasticidin for 9 days...18007609. Addgene Materials Used 1. lentiCas9-Blast (Plasmid #52962) was a gift from Feng Zhang and is described...BRDN0000561167) (Plasmid #80034) and BRAF gRNAs (BRDN0001145345 and BRDN0001146266) (Plasmid #76479 and #76480...transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then used ...expression in A549 cells transduced with lentiCas9-blast. Anti-Cas9 Western blotting was performed on whole-cell...then expanded for an additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed...viruses, but you can find a list of validated gRNA plasmids on our website. There are a few reasons to target...
Showing: 281 - 300 of 1034 results