We narrowed to 816 results for: LAS
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TypeBlog PostPublishedDec. 7, 2020, 2:53 p.m.... 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...
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The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog PostPublishedAug. 20, 2020, 2:30 p.m....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... -
Pushing the Limits of DNA Assembly
TypeBlog PostPublishedSept. 6, 2017, 1:01 p.m.... 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 ... -
Viral Vectors 101: Integrase-Deficient Lentiviral Vectors
TypeBlog PostPublishedMay 13, 2025, 1:15 p.m....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... -
CRISPR 101: Homology Directed Repair
TypeBlog PostPublishedJan. 26, 2023, 2:15 p.m....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... -
Antibodies 101: Isotypes
TypeBlog PostPublishedOct. 19, 2021, 1:15 p.m....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: ... -
Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs
TypeBlog PostPublishedJune 2, 2015, 6:06 p.m....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... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog PostPublishedJan. 31, 2019, 1:32 p.m....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 ... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog PostPublishedAug. 9, 2022, 1:15 p.m....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... -
Split-BioID: An Improved Method for Studying Protein-Protein Interactions
TypeBlog PostPublishedMay 1, 2018, 1:57 p.m....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...