We narrowed to 584 results for: abo.1
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TypeBlog PostPublishedJan. 8, 2019, 1:54 p.m....s pLKO.1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...experiments. Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David ...collection Find plasmids for your research Learn about our viral service ...
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Hot Plasmids - February 2022
TypeBlog PostPublishedFeb. 22, 2022, 2:15 p.m....include mEGFP, mCherry, and HaloTag. Figure 1: (a) Anti-FLAG frankenbody (FB) construction from ...Comm. 2019. https://doi.org/10.1038/s41467-019-10846-1. Optimized genetic screening technique using Cas12a... to Cas9’s larger expression requirements (Figure 1), but Cas12a has generally been underutilized in genetic... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here. ... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog PostPublishedAug. 15, 2019, 12:30 p.m.... requires selection. These two steps are (Figure 1): The initial plasmid integration step (the so-called...antibiotic resistance marker from the vector (abR, Figure 1). The resolution step (“double crossover”) which leads... loss of a specific phenotype (using sacB, Figure 1, further discussed below). Counter-selectable marker...Current protocols in molecular biology 106.1 (2014): 1-16. PubMed PMID: 24733238. Additional resources on... a sucrase that converts sucrose into a toxic metabolite. Cells that have undergone a double crossover...confirmation of successful single crossover – after about 48 hours, the resulting colonies turn subtly blue...blog posts Find our CRISPR blog posts Learn more about Lambda Red recombineering Resources on Addgene.... -
Plasmids 101: CcdB - The Toxic Key to Efficient Cloning
TypeBlog PostPublishedDec. 8, 2016, 3:30 p.m....succumb to the toxicity of CcdB. Figure 1: Image from Bernard, P., et al. showing the pKIL 18...submit a guest blog entry of your own! References 1. Bernard, P. “Positive Selection of Recombinant DNA... plasmid ccdB killer gene”. Gene. 1994 Oct 11;148(1):71-4. PubMed PMID: 7926841. 4. Bernard, P., et al...in modern cloning, and why should you learn more about it? Read on to find out how ccdB can make your cloning...of this system for enhancing cloning efficiency about 20 years ago and developed cloning vectors to harness... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog PostPublishedMay 11, 2021, 1:15 p.m....were found on the plasma membrane (top panel in Fig. 1). Of course, this was problematic, because the DREADD...plasma membranes in both species (bottom panel in Fig. 1). Our results indicate that the tag protein may have...protein of interest the membrane. Figure 1: Electron microscope images showing ultrastructural...Fluorescent 101 blog posts Read about chemogenetics vs. optogenetics Find more about the many types of protein... -
Michael Koeris' Journey from Grad Student to Entrepreneur: The Story of Sample 6
TypeBlog PostPublishedJan. 21, 2016, 4:52 p.m....Podcast breakdown (Time : Topic) 0:00 - 1:25 : Introduction 1:25 - 6:19 : Dr. Koeris' graduate work in...name from Novophage to Sample 6. To learn more about Novophage's successful transformation into Sample...to focus on a technology that one is passionate about developing? 38:00 - End : Dr. Koeris' role on the... -
Open Resources and Plasmid Tools For Studying C. elegans
TypeBlog PostPublishedJuly 18, 2019, 12:55 p.m....Zhang et al., 2015). In this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin...splitting cGAL4 in two and binding each half to a gp41-1-N-intein. Split inteins are protein domains that naturally...mutations with unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust...resources on the Addgene blog Learn more about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids...always emphasized the need for open science and collaboration. The field already has comprehensive reference...planned for August 15th, 2019. For more information about the aims and approach of CeNGEN, check out their...elegans. This database provides detailed information about C. elegans 3’UTRs structures, alternative polyadenylation... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog PostPublishedAug. 9, 2022, 1:15 p.m....unlimited supply of identical antibodies (See Fig 1.). rAbs are a relatively new technology that takes...conveniently modified and engineered. Fig. 1: Antibody production methods NeuroMab and others... the signal. If the exposure time is long (e.g. > 1/7s) then the signal is weak and vice versa for a strong...Have you ever been stuck googling about an antibody that will be essential to your new research project...high-quality antibodies for IHC. Working with collaborators and support from the Brain Research Through ...extensive library of monoclonal antibodies; Addgene collaborates with these companies to create and provide general...pattern should align with what the atlases (mentioned above) and literature suggest, providing an expectation... -
Management for Scientists: Seeking Feedback
TypeBlog PostPublishedJan. 8, 2015, 2:34 p.m....-public settings. Ask over coffee. Have regular 1:1 meetings with team members (don’t become known for...post, a study coming out of MITs Human Dynamics Laboratory identified five characteristics of very successful...conversations within the team. We are not talking about gossip here – this means encouraging a culture of...Medical School researcher Connie Cepko to learn about her management style. Seeking feedback frequently...information to make change whenever possible. Be public about changes influenced by feedback. If you don't do ... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog PostPublishedMarch 12, 2020, 1:15 p.m....used four different design strategies: Strategy #1 (ProA): Create promoters with sequences upstream of...active in retinal cell types in mice. Figure 1: The four strategies used to design synthetic promoters...highest rate of targeted expression. Surprisingly, <1% of synthetic promoters replicated the expression ...Addgene blog Read more about using AAV for retinal gene therapy Find more about precise neuronal targeting...injected into macaques and postmortem human retinas. About 25% of these AAV synthetic promoters successfully... -
Adeno Associated Virus (AAV) for Cell and Gene Therapy
TypeBlog PostPublishedNov. 7, 2017, 1:59 p.m....particularly passionate about HIV cure research and science communication. References 1. Gaspar, H. B., S...reconstitution with a normally functioning T cell repertoire (1). In this example, it was possible to transduce the...significant improvements in vision over a period of 1 to 3 years – the treatment appears to be both safe...Vectors Featured Topic Page Learn about CRISPR-AAV vectors Learn about Addgene AAV Quality Control Process... -
Genome-wide Screening Using CRISPR
TypeBlog PostPublishedAug. 18, 2015, 2:30 p.m...., check out Addgene's CRISPR Guide. Figure 1: Like this jar of jellybeans, a pooled CRISPR library... selecting a CRISPR library for your experiments. 1. What species are your cells derived from? Currently...a different gene. Libraries may be available in a 1-plasmid system, in which Cas9 is included on the gRNA-containing...preparation from Addgene, you can skip the steps above! Cells are then transduced with lentivirus containing...practices, CRISPR libraries can help you learn a lot about your phenotype of interest. For more detailed information...Libraries” (McDade et al., 2016). To learn a bit about how CRISPR/Cas9 can be used in other types of experiments... -
Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools
TypeBlog PostPublishedApril 23, 2018, 2 p.m....difficult-to-manipulate primary cells, achieving knockdown within 1-2 hr of reagent delivery. TRIM-Away is suitable for... inducer levels. That is, if transcription factor 1 controls expression of transcription factor 2, which...modulating the induction of transcription factors 1 and 2. See Aranda-Díaz et al. for more information...articles from April 2018. If you'd ever like to write about a recent plasmid deposit please sign up here. No... -
Lighting Up Cell Signaling with Photoswitchable Kinases
TypeBlog PostPublishedJuly 25, 2017, 1:20 p.m....current state (Green when off, dark when on). Figure 1: Photoswitchable MEK1 (psMEK1). Is it possible to ...Below are a few examples demonstrated in Zhou et al. 1. Testing kinase inhibitors Zhou et al examined the...the protein and pdDronpa are preserved. References 1. Zhou, Xin X., Linlin Z. Fan, Pengpeng Li, Kang Shen...Additional Resources on the Addgene Blog Learn More about Photoactivatable Fluorescent Proteins Control Protein... -
Choosing the B(right)est Fluorescent Protein: Photostability
TypeBlog PostPublishedJune 8, 2017, 2:30 p.m....various cyan fluorescent proteins is depicted in figure 1. Of note, the photobleaching rate does not depend ...to be used with the fluorescent protein. In figure 1 we provide an example of such a comparison between...the variants is comparable (lower panel of figure 1). To avoid changes in excitation power, it is important...Fluorescent Proteins for Multicolor Imaging Learn about Fluorescent Protein Biosensors Additional Resources... -
Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening
TypeBlog PostPublishedApril 12, 2016, 2:30 p.m...., including those found at Addgene! Figure 1: Strategy for assembling the barcoded gRNA library...pool-clone them into the pAWp28 storage vector (Figure 1): Forward oligo: 5’- CACCGNNNNNNNNNNNNNNNNNNNGTTTGGGTCTTCGAGAAGACCTATTCXXXXXXXXC... He can be contacted at [email protected]. References 1. Cheng, Allen A., Huiming Ding, and Timothy K. Lu....How Barcodes Can Be Used in Lineage Tracing Read about CRISPR/Optogentics Tools Resources on Addgene.org... -
The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years
TypeBlog PostPublishedJan. 8, 2014, 3:10 p.m....backbones for shRNA expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids ...years of plasmid sharing Interested in more facts about Addgene's 10 years of plasmid sharing? Check out... -
Progress Towards a PAM-Free CRISPR
TypeBlog PostPublishedMarch 7, 2024, 2:39 p.m....nucleotides are represented equally, that’s only a 1/16 chance of having a PAM at a given site. While it... NYN/NRN NYN is less effective than NRN Table 1 - Summary of PAM-flexible Cas enzymes Are we truly...editable with CRISPR technology. Fig. 1: SpCas9 has sparse PAM coverage in the human genome...experiments, therefore, you often must make a choice about what is your top priority: location or efficiency...These Cas proteins have everything we know and love about Cas9, but with broader PAM compatibility. This alternative... -
Plasmids 101: Terminators and PolyA signals
TypeBlog PostPublishedMarch 31, 2016, 2:30 p.m....initiating cleavage of the transcript. Figure 1: A predicted conserved secondary structure and sequence...prokaryotic terminators and deposited several with Addgene (1). Prokaryotic polyadenylation Although mostly thought...constitutive RNA transport element lead to enhanced HIV-1 Gag expression in a context-dependent manner. Nucleic...additional helper sequences (2-3). As alluded to above, termination and polyadenylation of Polymerase II...how the Promoter Controls Gene Expression Learn about New Cloning Techniques Read all of our Plasmids ... -
Plasmids 101: Repressible Promoters
TypeBlog PostPublishedDec. 20, 2022, 2:15 p.m....Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018...and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed Central...Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939-6371-3_3. PMID: 27730575; PMCID: PMC5270762. ... and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed PMID: 6337371. PubMed Central...can be used in the repressible manner described above, as well as the inducible manner detailed in our...necessary to have all 3 of the different binary systems above if they all behave very similarly?” The simple answer...