...that they called a Glycosylase Base Editor, or GBE, for its reliance on the glycosylase UNG to crease abasic...Converts C → G; also called a glycosylase base editor (GBE). GBE Glycosylase Base Editor A base editor ...Uracil DNA N-Glycosylase Removes uracil from DNA, creating an abasic site. UGI Uracil Glycosylase Inhibitor...at its original job. They removed the uracil glycosylase inhibitor (UGI) components, which inhibit BER...instead introduced its opposite, a uracil DNA N-glycosylase (UNG) from E. coli. The resulting construct, ..., Rosser, S. J., Bi, C., & Zhang, X. (2021). Glycosylase base editors enable C-to-A and C-to-G base changes...Additional resources on Addgene.org Browse CRISPR plasmids for base editing Get a broad overview with our...

...Biotechnology which allows researchers to deposit plasmids containing the fluorescent protein mNeonGreen....with other fluorescent proteins. Find mNeonGreen plasmids here! mTFP1: a monomeric teal fluorescent protein...492 nm, respectively), they gave this a teal classification rather than cyan and named the protein mTFP1...physiologically relevant changes in pH. Find mTFP1 plasmids here! mWasabi: a bright green fluorescent protein...excite at 400 nm while EGFP does. Find mWasabi plasmids here! One of the most notable things about these...are best suited for certain studies. If you have plasmids containing mNeonGreen, mTFP1, or mWasabi that ...with the scientific community, please deposit the plasmids at Addgene! References Ai, Hui-wang, et al. "...

...More than a model System - Arabidopsis in the classroom With funding from the American Society of Plant... the kits, known collectively as Greening the Classroom, were developed and tested by ABRC staff. These...an ordering account. Using Arabidopsis in the classroom has many benefits. Engaging students in active...science curriculum. Growing Arabidopsis in the classroom does not require a huge investment in resources.... Another benefit of using Arabidopsis in the classroom is that staff from the ABRC are here to support... people in science learning. Whether visiting classrooms, hosting field trips, conducting teacher workshops...resource or you want to introduce Arabidopsis to your class for the first time – we want to hear from you. Reach...

...About Other Hot Plasmid Technologies Read Other Synthetic Biology Posts  Suggest Plasmid Tools for the ...Resources on Addgene.org Read our Hot Plasmids Articles Find Plasmids for Bacterial Expression ...to changing pH, and they can rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS...these fluorescent R bodies can rupture E. coli spheroplasts (cells which have had their cell walls removed...produced R-bodies that could extend at different pHs. Plasmids expressing expressing both wild type and mutant...acssynbio.5b00237. PubMed PMID: 26814170. Find Plasmids from This Publication at Addgene 4. Petroni, ...

...the ability to distribute plasmids with the piggyBac™ transposon. These plasmids, when combined with a source...useful resource. While Addgene cannot distribute plasmids with the piggyBac™ transposase itself, please ...genome fully intact, while, with a standard DNA plasmid transfection, a double-stranded break may occur...by utilizing a piggyBac™ transposon in a donor plasmid carrying the DNA edit of interest, along with a...piggyBac™ transposase enzyme [4]. By using a donor plasmid containing a negative selection marker such as ...kill off any cells that still contain the donor plasmid after excision by PBx, thereby leaving behind only...cytotoxicity [5]. Second, instead of using a donor-plasmid, you could use a single-stranded oligodeoxynucleotide...

...and to make the desired genome edit:  A donor plasmid that contains a puromycin resistance gene linked...the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The Madison lab also created...in the reporter gene). A control RFP expression plasmid.  With this set up, all successfully transfected... and repaired via HDR using the supplied donor plasmid will express GFP. If there’s GFP expression, presumably...through the presence of a fluorescent reporter. Lastly, once a mutant is identified, the PB transposase...just your desired genomic edit.  Find SRIRACCHA plasmids at Addgene GEMCherry2, an improved monomeric fluorescent...embryonic stem cells.  Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter...

...ligand-bound GPCRs of interest. The TRUPATH plasmid kit consists of 20 plasmids encoding 14 alpha, 2 beta, and 4 ...response? Let’s take a look at a few examples with plasmids deposited at Addgene.  PRESTO-TANGO: The ligands...Bryan Roth’s lab also assembled a collection of plasmids to elucidate the role of 14 of the 16 non-visual...pathways mediated by GPCRs? The Kruse Lab constructed plasmids that can be used in peroxidase-catalyzed proximity...inhibit neuronal firing. As GPCRs are the largest class of signal transducing receptors in the brain they... (firing or inhibition). AAV preps from DREADD plasmids allow you to easily express a DREADD receptor ...Addgene's chemogenetics guide Find chemogenetics plasmids for your research ...

...Standards Find Plasmids for Creating and Controlling Gene Networks Browse All SynBio Plasmids ... of “bringing tomorrow’s science into today’s classrooms,” BioBuilder answers the call by offering a variety...forget the technical details they hear in their classes,” she said, “but they remember the moments when..., Kuldell also knew that the best days in the classroom are characterized by intense student engagement...and how — some choose to run full semester-long classes, some pick and choose a few lessons to include ...book chapter and an illustrated, downloadable classroom poster — with both of these resources road tested...any stage of completion, from pitch to prototype. Last year’s BioBuilderClub teams’ biological engineering...

...is a global nonprofit plasmid repository. Over 2,000 labs have deposited plasmids to Addgene and we distribute...opportunity for Addgene to present our own data on plasmid deposits and distirbution for these fast moving...distribute over 130,000 plasmids in 2014. Thus, we are in a unique position to observe and quantify how...scientific community. Data on CRISPR and synBio plasmid sharing In a recent review on CRISPR technology...trends for Zinc Finger, TALE, CRISPR, and SynBio plasmids over time. You can see these data in the slide...molecular biology technology? Have you deposited your plasmids yet?      ...

...DEPC.  Ensure that plasticware or glassware used is RNase free. RNase-free plasticware is readily available...doing something without a kit has advantages: less plastic waste, less expense, and less of being left with...available from scientific suppliers and glassware should be treated with a DEPC solution for 1 hour, and autoclaved...autoclaved to remove residual DEPC. Alternatively, glassware can be baked at 180°C for at least 4 hours.   ...selectively precipitate RNA, but not DNA and proteins. Lastly, using a DNase (there are several DNase enzyme ...on the Addgene blog Read blog posts related to plasmid protocols Find protocols for purifying DNA without...

...fiberoptic cannulas directly implanted in the injection site and connected to a laser. The laser flashes light... about FLEx vectors Download our Plasmids 101 eBook Read our plasmids blog posts Resources on the Addgene...molecular biology reference Find Cre-lox plasmids at Addgene Browse plasmids at Addgene ...received her PhD in Virology from the University of Glasgow. She then moved to France where she worked as a...

... scientists share plasmids across the globe, we’re working on ways to send plasmids as DNA as opposed ...fiIling The majority of Addgene plasmids are shipped as agar stabs in glass vials. Have you ever wondered... transformation. With our ever growing rate of plasmid deposits (over 100 deposits per month!), manual...Qiagen BIOROBOT Universal Platform and it suits our plasmid prepping needs perfectly. Working with Qiagen, ...used to be a lab assistant, but with increased plasmid demand it was too time consuming to complete this...There is a tube that goes from the needle to the flask with agar in it. The agar sits on a hot plate with...

...up your or your class’s streak plate skills? Find the chromoprotein expressing plasmids here, and check... Find fluorescent protein plasmids at Addgene Find chromoprotein plasmids at Addgene Check out additional...other species.  When he found the chromoprotein plasmids available from Addgene, he knew it would be a ...hopefully revealing single, isolated colonies in the last quadrant after incubation. The challenge is usually...molecular biology. “What I'd like to do with the plasmids from Addgene is have the students transform them...

...Gonçalves and Ventura labs’ plasmids, including protocols, check out the plasmids at Addgene: pAdSh.PGK.Cas9...looking for a broader range of CRISPRs plasmid tools, find more plasmids, CRISPR technology guides, FAQs, and...Originally published Sep 30, 2014 and last updated Dec 10, 2020 by Benoit Giquel.   Adenoviral vectors...description of their experimental protocol. The three plasmids which have been deposited to Addgene are: pAdSh.PGK.Cas9...for instance in inherited disorders such as B-thalassemia, Sickle Cells Disease, Duchenne Muscular Dystrophy...nonalcoholic steatohepatitis cancer. In deficiencies of plasma proteins where Adv vector delivery is often used...

...Fluorescent Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...for a transcriptional reporter or as a general cytoplasmic label of cells, there's no problem. You're OK...tag with a serious drawback. The standard EGFP plasmid that used to be sold by Clontech and is in a freezer...stacked structures including organized smooth endoplasmic reticulum (OSER) whorls. 2)A dimerizing protein...dramatic. For a membrane POI localized to the endoplasmic reticulum, the normally spiderweb-like pattern...dense stacked membranes termed Organized Smooth Endoplasmic Reticulum or OSER if a dimerizing FP is fused...quality control of secretory proteins in the Endoplasmic Reticulum and live cell imaging approaches. Erik...

...2011). But this requires other lasers than the standard Ti:Sapphire laser. It is also important to finely...Two low-energy photons (typically from the same laser) cooperate to cause a higher-energy electronic transition...two-photon probes when using a standard Ti:Sapphire Laser include: Blue-green fluorescent proteins like EGFP...fluorescent proteins are excited by 750 nm to 760 nm laser light, enabling dual color imaging studies with ...researcher can flank fluorescent proteins in a plasmid with loxP sites such that their expression is turned... the brains of mice chronically infected with Toxoplasma gondii." The Journal of Immunology 182.10 (2009...cells and dendritic cells during infection with Toxoplasma gondii." PLoS Pathog 5.7 (2009): e1000505. PubMed...

...Resources on Addgene.org Find Plasmids for FRET Browse Empty Plasmids to Fluorescent Protein Fusions ...oocytes, mainly). Nevertheless, tRNA suppression plasmids are available from depositing labs that have mastered...the potential rewards could be substantial: Plasmid tRNA Synthetase Expression Host pMAH-POLY Polyspecific...Fig. 1) which can be carried out using Addgene plasmids. His-tag labeling reagents You may already have...cell membranes, so they are best used either with plasma membrane proteins (such as GPCRs, ion channels,...40-72 Intact Cells Cy3NTAc Poly His  44 35-63 Plasma membrane protein, permeabilized cells  Cy5NTAc...38-70 Intact Cells Cy3NTAc Poly His  63 50-90 Plasma membrane protein, permeabilized cells  Cy5NTAc...

...suppressed and NES activity is enhanced, leading to cytoplasmic translocation of the reporter. As a result, the...phosphorylation status. The relative ratio of cytoplasmic vs. nuclear fluorescence serves as a proxy for...monitor global kinase activity that occurs in the cytoplasm and/or nucleus, while FRET-based reporters can...because KTRs shuttle between the nucleus and the cytoplasm, nuclear export and import rates can influence...ERK, or a backbone to use for custom KTR design, plasmids are available from Addgene! References Kudo, ...protocols, 13 1, 155-169. PMID: 29266096. Find the KTR plasmids here! Additional Resources on the Addgene Blog...Additional Resources on Addgene.org Find FRET plasmids Find fluorescent biosensors ...

...transformation Read our Plasmids 101 blog posts Resources on Addgene.org Browse our Plant Plasmids and Resources...page Find synthetic biology plant plasmids Find CRISPR plant plasmids ... the production of cancer therapeutic drugs, vinblastine and vincristine, are regulated in the medicinal...cannot be studied in hairy roots, as functional chloroplasts and leaf specific cell types are missing in ...its function. In nature Agrobacterium contains a plasmid with a T-DNA (transfer-DNA) region. During infection...localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different leaf...

...days, we would ‘spot-check’ critical regions of plasmids deposited with us via Sanger sequencing, while...while more recently we have moved to full plasmid sequencing at our facility using Illumina MiSeq Next Generation...lab work as we grew. As the number of deposited plasmids increased, the accompanying sequencing data collection...transform the raw sequencing data into a complete plasmid sequence that would be ready for our QC team to...With our outline of individual steps to automate plasmid sequence assembly, and possible solutions for each...up with our sequencing instrument (hundreds of plasmids each week!), we already planned to use cloud computing...computer. Eventually, our workflow would handle plasmid sequences on a 96-well plate basis, but for rapid...