...and tech transfer communities who help guide and promote Addgene in its mission to make research easier.... interesting biological questions, you need a tremendous amount of technology development alongside it...what is worth having. The work that [Addgene’s] outreach scientists do to comb through the literature, ...

...the Francis Crick Institute concluded “Aspirin promotes autophagy, inhibits platelet aggregation, and ... that fasting has a positive effect on cancer treatment (Ref 1). As described in the presentation, starvation...favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive reaction...

...visualization and expressed from a heart-specific promoter. Fish were labeled with MG-2I, either by incubating...hearts 24 hours post treatment and no mCer3 cells present 96 hours post treatment (see figure 2). Induced...culture experiments, only dL5**-expressing cells treated with MG-2I dye and exposed to light stained dead...Chromophore-assisted laser inactivation in cell biology. Trends in Cell Biology,18(9), 443-450. doi:10.1016/j.tcb...

...assembly reaction. DNA fragments with the desired promoter, selectable marker, and target gene, for example...and improvements in these methodologies offer streamlined and simplified alternatives to traditional cloning...

...Expression is driven by a fairly strong EF1alpha promoter. We have verified expression of correctly-sized...straight-forward. All constructs are tagged with a 2xStrep-tag for easy detection and affinity purification...

...studying how that one protein interacts with that one promoter can tell you, it can be easy to get lost in the...Fight Rare disease with iPSCs Read About Addgene's Outreach Efforts in the Microbiology Community Sign Up ...

...changes were introduced to nCas9. These changes promote nCas9’s dissociation from DNA after nicking and...bacteria resistant to both spectinomycin and streptomycin, but could also be used for studying epistatic...

...Community (MCC) Collection Pleiades Promoter Project: mini-promoters to drive selective expression in the...Syndrome Plasmid Collection and Resource Center TREAT-AD Plasmid Collection Visualization Michael Davidson...

...recognition of transcription-associated DNA damage that promotes homologous recombination. Gong F, Chiu LY, Cox...nucleosomes, loops, chromosomal domains, or how stretches of DNA on different chromosomes interact with ...K, Wang W, Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5):625-36. An architectural...

...Viral expression cassettes typically contain a promoter (inducible or constitutive), your gene of interest...the last decade, viral applications in cancer treatment and gene therapy have also been in the limelight...approved the first oncolytic viral therapy in 2015 to treat melanoma, and many more are in the pipeline. Viruses...

... 2008). Obligate dimers (such as dTomato) that promote interactions between the FPs interfere with native...Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced microscopy, University of Amsterdam...applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications...Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced microscopy (University of Amsterdam...

...-loops described above play distinct roles in promoting cellular processes and functions, but they also...production. In the same vein, genotoxic stress, such as chemotherapy treatment or ionizing radiation, can also...

...complex, to temporarily inhibit this process and promote copying of the edited strand to the unedited strand... co-express mismatch repair inhibitor MLH1dn Promote desired resolution of heteroduplex     ✓ ...non-homologous end joining. As a result, Cas9 treatment causes most products to be indels while the efficiency... In addition to the numerous innovations that streamline the process of prime editing, the Liu lab contributed...efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime editor; ... 2023) Prime editing has a lot of promise for treating genetic diseases. In our summer Hot Plasmids post...

...several weeks which may also depend on transgene or promoter choice. Behaviors you can study using viral vectors...into the experimental setup, the mouse becomes stressed. It will likely move around faster in the following...some time to adapt to a testing apparatus. General stress levels of the test animal may also be reduced by...noise should be avoided as this will also increase stress levels and affect behavior. A fact to consider:... groups), health status, diet, cage enrichment, stress level, origin (vendor or in-house mouse breeding...

...recombinase from either a heat-shock promoter or a tissue-specific promoter and expression of the target FLP-out... from either a ubiquitous or a tissue-specific promoter. cGAL and Split cGAL plasmids - Paul Sternberg...Jorgensen Lab. A collection of plasmids that streamlines the production of targeting vectors for tag insertion...

...nuclear localization, binding to the appropriate promoter sequence and activation of gene expression. In...bulk cell experiments, Clift et al. observed no stress or damage upon TRIM-Away, and the electroporated...re-adhered to the culture plate within 4 hr of treatment. In some primary cell lines, endogenous TRIM21...Listen to the Fire-Cas9 Podcast Segment Jerry Crabtree’s lab has developed a new CRISPR/Cas9-based system...

...sequence! In bacteria, Cas13d-mediated cleavage promotes collateral cleavage of other RNAs. As with other...displays very high on-target efficiency; where shRNA treatment produced 500-900 significant off-targets, CasRx...

...science and engineering. “Fix the Institutions” promotes gender equality in careers through structural ...factors may include cell type, the disease being treated, and other hormonal, immunological, and environmental...

...binding proteins with Cas9 guided by different promoters. The challenge in building genetic circuits is...anywhere else, but for synthetic circuits, they are extremely orthogonal. It’s a different perspective and you...

...dopamine is an important neurotransmitter that promotes reward-motivated behavior, if the channelrhodopsins...as predict the channelrhodopsin’s photocurrent strength, wavelength sensitivity, and off-kinetics.   This...did not turn. Left-turning behavior in ChRger2-treated mice ceased when the light was turned off.  Together...

...sgRNA with U6 promoter 48962 Worm HindIII none S. pyogenes de Bono sgRNA with rpr-1 promoter 48961 Worm ...of two sgRNAs from independent U6 promoters and Cas9 from CBh promoter. Ventura A CRISPR/Cas9 toolkit for...along with 1-4 sgRNAs expressed from independent promoters. sgRNAs are cloned into separate vectors and then... assembly of single gRNA vectors with various promoters, plasmids for assembling multiple gRNAs into one...pGGDestTol2LC vector. Uses five distinct zebrafish U6 promoters for sgRNA expression. Chen pCFD4-U6:1_U6:3tandemgRNAs...expression of two gRNAs from Drosophila U6:1 and U6:3 promoters. Bullock pCFD5 Drosophila Utilizes endogenous ...Zhang Cas9/pTREX-n 68708 Other/Trypanosoma cruzi yes, cut S. pyogenes Neo Docampo tdTomato/pTREX-b 68709 ...

...gene expression and the selection of specific promoters may also help AAV vectors evade host immunity....potentially be treated by correcting the mutation or replacing the gene. A notable example is the treatment of Severe...conditions, opening up new avenues for therapeutics that treat disease at the molecular level. For instance, if...function. To date, gene therapy has been used to treat 10 infants with this disease. To do so, their T-...childhood onset of blindness and was thought to be untreatable until 2008. Following a single dose of subretinal...in vision over a period of 1 to 3 years – the treatment appears to be both safe and effective. Challenges...antibodies that neutralize the AAV vectors used to treat them. Scientists are working to overcome this issue...

...discussions. Get your voice out there. Use Twitter to promote your other science communication activities, e....engagement - Participate in science festivals and outreach If you are comfortable talking in public, volunteer...groups, charities and NGOs are often looking for outreach volunteers as well. 4) Blog - Hone your writing... a blog is not only a great form of scientific outreach, it also helps you to hone your writing skills...science communication. The National Co-Ordinating Centre for Public Engagement (NCCPE) provides on-site ...

...incorporating it into a fourth plasmid. In addition, the promoter of the 5’ long terminal repeat (LTR) is mutated...experience of end-users, size of the transgene, and downstream applications should all be considered when choosing...factors such as resources available, cost and downstream applications. For example, while the PEG precipitation...be the best method when pure virus is required downstream. When assessing lentiviral titer, don't compare...

...help them navigate the scientific enterprise and promote their voices in driving changes in science. Follow...others in Boston (some of them literally across the street from these institutions) are raising salaries. ...

...its paralog RC2H2 are RNA binding proteins that promote RNA decay and their upregulation is correlated ...little was one minute BASU generated a strong streptavidin signal (~250-fold enrichment) when compared ...

...Program at Harvard Law School. He has continually promoted the cause of increasing data collection about ...Symposium Boston 2015 you can register here or live stream the event on October 22nd and 23rd here. Many ...

...orientation. This allows for multiple DNA components (promoters, genes, terminators, etc) to be assembled in the...ligation process is close to 100% efficient. Another strength of Golden Gate cloning is its scalability. Unique...

...under the control of Murine Stem Cell retrovirus promoter for mammalian gene knockdown. Vector provided ...sequencing to perform multiplexed analysis of cells pre-treated with a compound of interest. Lentiviral vectors...

... had multiple integration events. In addition, promoter choice can also impact transgene expression. The...transduced with serial dilutions of a viral prep, treated with the antibiotic, and colony-forming units quantified...

...Proximal Promoter Further upstream from the core promoter you will find the proximal promoter which contains...Reference Molecular Biology Reference Promoters Promoters Definition A promoter is a region of DNA where transcription...silencers. Promoter Regions There are three main portions that make up a promoter: core promoter, proximal...bind. Distal Promoter The final portion of the promoter region is called the distal promoter which is upstream...mammalian promoter PGK Constitutive Mammalian promoter from phospholycerate kinase gene TRE Inducible ...Specific Drosophila promoter containing Gal4 binding sites Bacterial Promoters Promoters in bacteria contain...tryptophan Promoter from E. coli tryptophan operon Ptac Regulated like the lac promoter Hybrid promoter of lac...

...surprise since the PAM flexibility could in theory promote off-target binding. The activity of SpRY was validated...precise targeting (give or take a bp or two upstream or downstream) for most gene targeting experiments. Between...

...decreases. Hypermethylation of CpG islands in the promoter region of tumor suppressor genes can repress their...study and treat complex or multifactorial diseases, enabling faster diagnosis and treatment and improving...

...universities stressful or simply not feasible. Through the virtual systems created and promoted during COVID...fit for you without the financial and logistical stress that accompanies traveling for campus visits. This...have to worry about traveling during an already stressful time, and I was more relaxed during both sets ...their end. Though the quick changes in plans was stressful in and of itself, the flexibility allowed through...using the virtual world to your advantage can strengthen your application and help you decide if a program...

...Yannick Doyon 214916 TRE-TDP-43ΔNLS-mClover3 TARDBP mClover3 TRE ALS Ophir Shalem 214917 TRE-mClover3-TDP-43ΔNLS...Yeast Other Promoter CMV T7 polH GAL Other Clear Filters ID Plasmid Name Gene Tags Promoter Disease PI ...pEHA1628) SNCA Avi TRE Parkinson's Vikram Khurana 210360 TMEM106B-2Strep TMEM106B Strep CMV, T7 Frontotemporal...Flag CMV ALS Yossi Shiloh 34927 pTRE_tau-LacZ::tTA-H100Y MAPT LacZ TRE Parkinson's, FTD Mark Mayford 35000...telangiectasia Stephen Elledge 43918 tetO-ALN NR4A2 His, V5 TRE Parkinson's John Gearhart 44333 pMX_mTBP TBP LTR ...Douglas Green 50487 pSLIK-NFLAG-hFUS FUS Flag, His TRE ALS Zissimos Mourelatos 51146 DsRed-p150 217-548 ...Dent 188572 Tet-off TauRD (P301L/V337M) MAPT Myc TRE Parkinson's, FTD Franz-Ulrich Hartl 188573 Tet-off...

...complexed with a gRNA that directs them to the promoter of a gene of interest, they have been shown to...conventionally used SpCas9 (derived from the bacterium Streptococcus pyogenes). Consequently, fewer genetic tools...complex. When this complex is directed to a target upstream of the transcriptional start site (TSS) of a gene...competent to cut DNA. When directed to a site downstream of the transcriptional start site of a gene of...

... al. YY1 Is a Structural Regulator of Enhancer-Promoter Loops. Cell. December 14 2017. 171(7):1573-1588.... Test dTAG molecule toxicity in new cell lines Treat parental cells (or cells expressing a control --...

...plasmid harbors a multi-cloning site flanked by T7 promoters to allow for bidirectional transcription to create...to the worms. The phenotypes of RNAi knockdown treatment should be apparent after three dsRNA feedings....

...gene expression in C.elegans. In this system a promoter controls the expression of a GAL-4 driver that...that can bind to activation sequences upstream of genes to stimulate expression (Wang et al., 2016). In ...

...express hybrid Cas9-Cas12a gRNAs under a single U6 promoter. The paralog and dual-targeting hybrid gRNA library...Addgene Sc++ is a codon-optimized, engineered Streptococcus canis Cas9 nuclease that has a broad 5′-NNG-...

...functional role of DNA demethylation at specific promoters by targeted steric blockage of DNA methyltransferase...understand mechanisms and, eventually, develop treatments, we need tools to specifically edit methylation...

...activity Regulating Acr proteins by inducible promoters, light, or small molecules could achieve rapid...meningitidis (Nme) (Pawluk et al., 2016). While Streptococcus pyogenes Cas9 (SpyCas9) is the most used and...system. 5. Phage therapy Phage therapy uses phage to treat bacterial infections as an alternative to antibiotics...

...walk into any supermarket now without seeing a promotion for antibiotic-free, farmed meats. We can focus...antibiotics. When we use antibiotics to treat infections, we are treating microbes with compounds similar to...will be more difficult to treat later. A similar situation can occur if you treat a bacterial infection with...more different classes, or types, of drugs used to treat them. Very often, this is what it means to be a ...often very difficult and very time consuming to treat. Learn How Antibiotic Resistance Genes Are Used ...antibiotics in livestock farming Lengthy courses of treatment with antibiotics These scenarios expose bacterial...the population and making them more difficult to treat in the future. Patients not finishing the whole...

...on the Addgene Blog Origin of Replication The Promoter Region – Let's Go! Browse All of Our Plasmids ...in cells treated with Plasmid A is similar to the expression level of Gene X in the untreated cells—but...be determined empirically.  Negative controls: untreated cells, empty vector control, and non-targeting...is observed). In any plasmid-based experiment, untreated cells should be included as these provide the ...

...Truncated MeCP2 promoter-driven SpCas9; for neuronal expression 60958 : PX552; U6 promoter-driven; for sgRNA...tyroxine binding globulin (TBG, PX602 [#61593] ) promoter, and a U6-driven single guide RNA. The vector ...codon-optimized SpCas9, driven by the truncated MeCP2 promoter (pAAV-pMecp2-SpCas9-spA) for expressing Cas9 in...FA, Hsu PD, Lin CY, Gootenberg JS, Konermann S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang...CRISPR-Cas9 complex. Konermann S*, Brigham MD*, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib...

...an enhanced pU6cm promoter, viral replicon amplification of cargo, and heat treatment.  Figure 1...dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage of the self-labeling tags ...Turner, E.D., Abdelhamid, M.A.S., Craggs, T.D., & Twelvetrees A.E. (2024) Kinesin-1 is highly flexible and ...

...shown in tables 1 and 2, with a cell-type specific promoter or enhancer now allows for cell-type-restricted...special CREATE cap plasmid, loxP sites flank the downstream polyA sequence. When viruses produced from this...site of the injection. IV injection into the bloodstream allows for dispersion of the virus but it has...applications (mechanistic studies and gene therapy treatments for neurological diseases). The Gradinaru Lab...

...regulatory regions in the genome, including enhancers, promoters, insulators, and more. I then identified valid...and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning of the third chromosome... Let’s now examine how we can locate powerful stretches of DNA known as enhancers with the help of genome...light on dark matter Precise knowledge of which stretches of the genome are pillars for stem cells, or metastasized...

...plasmid name for ordering information. ID Name Promoter Description Category PI 157970 pTR-UF11 chimeric...of retinal bipolar cells with improved mGluR6 promoters in rodents and primates. Mol Ther . 2016 Aug;23...by the physiologic body fluids, such as blood, vitreous, CSF, etc. Not suitable as a buffer for vectors...enhanced transduction of the retina following intravitreal injection in mice and marmosets as compared ...enhanced transduction of the retina following intravitreal injection in mice and macaque. This serotype... citations include: Scalabrino, et al. 2015. Intravitreal delivery of a novel AAV vector targets ON bipolar...

...continuous outflow of phage, increasing stringency and promoting survival of only the broadest-targeting variants..., also has a strict editing window about 15 nt upstream of the PAM. Even if there’s a PAM near your insertion...