...AAV-pgk-Cre PGK Cre expression Recombinases Aebischer 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a Cre and physically-separate...Cre.SV40 CamKII Cre expression Recombinases Wilson 107738 pAAV-hSyn-Cre-P2A-dTomato Syn Cre expression with... by using this serotype to deliver Cre-dependent transgenes in Cre transgenic lines. As part of our standard...-EGFP Syn EGFP, Cre-dependent Control Roth 55650 pAAV-hSyn Con/Fon EYFP Syn EYFP, Cre and Flp-dependent...pAAV-FLEX-tdTomato CAG tdTomato, Cre-dependent Control Boyden 27056 pAAV-Ef1a-DIO EYFP EF1a EYFP, Cre-dependent Control...Expresses NLS-mCherry in the absence of Cre, and expresses NLS-EGFP in Cre-positive cells. Control Harvey 137164...expression Recombinases Deisseroth 55636 pAAV-EF1a-Cre EF1a Cre expression Recombinases Deisseroth 55637 pAAV-EF1a-Flpo...

... Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein Plasmids and Resources...already have created a transgenic GFP-containing organism,” says Hanson, “using a gene encoding one of the... Methods. 2009 Feb 6(2):131-3.  PubMed. Find plasmids from this publication at Addgene. Subach OM,...Resources Relevant Blog Posts Plasmids 101: Green Fluorescent Protein Which Fluorescent Protein Should I...characterized, it turns out this protein has a few more secrets - during a collaboration, members of Maureen Hanson...necessary to observe a signal. Red signal intensity increases with GFP concentration and saturates around 30...

...selectable with the antibiotic whose resistance gene is encoded in the viral vector. This antibiotic selection ...percentage of surviving cells may be the result of plasmid integration via recombination during transfection...production of virus. Alternatively, if the virus encodes a fluorescent protein, the packaging cells could... prior to their transduction has been shown to increase AAV transduction efficiency by approximately 15...efficiency depends on viral titer Both of these can be increased by concentrating a viral stock. Different titering... virus-cell contact A variety of reagents can increase transduction efficiency. These reagents typically...reagent that is commonly used and has been shown to increase transduction efficiency by 10-fold (Davis et al...

...are transfected with plasmids expressing the GPCR of interest, a Gα-Rluc8 plasmid, and the appropriate...downstream effects.  Bryan Roth’s lab created a collection of plasmids called TRUPATH to interrogate 14 of...their mechanism. These plasmids are available as a collection, facilitating screening across the transducerome...But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002) and many downstream...TRUPATH be used for? The Roth lab used the TRUPATH plasmids in a number of different experimental approaches...Addgene.org Browse the catalog of fluorescent protein plasmids Find biosensors for your experiments ...resulting in decreased BRET2 signal. With increased concentration of drug, the BRET2 signal decreases. ...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...melanogaster. Get the plasmids at Addgene The Cas Hybrid for Multiplexed Editing and screening Applications (...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...different neurotransmitters. Using genetically encoded fluorescent proteins coupled with Fibronectin intrabodies...targeting inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration...

...you find what you need.   More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular...need. You will also have to make sure that your plasmid has been incorporated into the host organism, usually...open reading frame (ORF) of interest –  try pRS plasmid shuttle vectors. You may prefer the more traditional...YGOI in a pattern that allows the function of the encoded protein to be assayed (e.g. pPD95_86 and pPD57_...details and guidance for choosing your perfect plasmid backbone. Or just ask –  leave a message in the...Molecular Cloning Guide & Other Plasmid Protocols List of common epitope tags ... bacteria and a mammalian selection cassette to screen for positive transfected cells. The addition of...

...spatial control. Optogenetics uses genetically encoded tools, such as microbial opsins, to control cellular...reporter gene in mice using Minicircle DNA vectors encoding the FAST constructs. This proof-of-concept model...approach. These researchers used a genetically encoded photocaging technique to insert a light-removable.... presented data showing that this genetically encoded, photocaged Cas9 system could silence endogenous...Website Browse Our Collection of Optogenetics Plasmids Check out Our Genome Engineering Resources Brush...researchers saw an 11-fold increase in mRNA levels after 2 hr and a 400-fold increase after 30 hr. The system...(CIB1) proteins. The goal of both groups was to create a system that would use light to turn on and off...

...Addgene.org Find plasmids for in vivo imaging with fluorescent proteins Browse luciferase plasmids Find fluorescent... Genes (ARGs) are engineered gene clusters that encode proteins that form bacterial gas vesicles. These...acoustic reporters for your research? arg1 and arg2 plasmids are available from Addgene! References 1 Bourdeau...operons from two different bacteria, Bourdeau et al created two constructs optimized for expression in E. coli... and have already developed a high-throughput screening method for serial acoustic collapse to aid in ...

...studying four of those rare diseases will receive plasmids from Addgene worth a total of $5000.  Lowe Syndrome...involving the primary cilia. They’ll put a variety of plasmids to work to explore the effect of various OCRL1...there. This will help us acquire a lot of those plasmids so that we can try many different things in parallel..., will use GFP-ATP1A3, FLAG-ATP1A3, and ATP1B3 plasmids to explore the functional differences among mutations...responsible for the disease as ATP1A3. The gene encodes a sodium-potassium pump expressed specifically ...mutations. More specifically, they’ve proposed to use plasmids with modified nickase Cas9 protein in order to...This recessive genetic condition leaves patients incredibly sensitive to sunlight and prone to developing...

...synthetic biology page Search for a plasmid Find Addgene's most popular plasmids ...been coming to the Addgene website in search of plasmids. Now, scientists are beginning to see Addgene ...Addgene as a large data set. Addgene has over 65,000 plasmids in the repository, each verified by sequencing...preferences for the codons they use; even if two species encode the same protein, their gene sequence may have ... recently published a tool (trained on Addgene plasmid datasets) to predict the lab-of-origin of engineered...series of simulations and machine learning, they created a classifier that predicts whether a gene was natural...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available... Xiao Wang Lab has recently deposited a set of plasmids that helps identify base edited cell populations...populations without end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter...available from Addgene, head over to our CRISPR Plasmids and Resources page. Here are some highlights from the...gene. The lab modified the lentiviral backbone to encode the CD90.1 selection marker, Thy1.1, in addition...

...experiment. We’ll start with ChIP-seq data from the ENCODE project, find enhancers in embryonic stem cells...sites from 125 different human cell types from the ENCODE Project. DHS sites are the most inclusive markers...Genome Edit Resources on Addgene.org Find CRISPR Plasmids for Your Research Check out Our CRISPR Guide Find...CRISPR screen to disrupt these enhancers. The third and final step is to conduct our CRISPR screen. We start... the marvel of DNA compaction, an enhancer can increase the expression of a gene that is millions of base...cells, and conclude with a targeted CRISPR screen that can disrupt these enhancers. A real-world experiment...that have NANOG binding sites 2. Design a CRISPR screen to target and disrupt the NANOG enhancers 3. CRISPR...

...more cells = more plasmid. Plasmids can vary in copy number, the number of plasmid copies supported within... optimize your plasmid prep We'll discuss the following techniques to increase plasmid yield. Keep in ...may lead to loss of the plasmid, and using older plates could increase plasmid loss or mutation. Let it...where your plasmid yield is sub-optimal? If you are consistently getting sub-optimal plasmid yields from...genotype, as these mutations improve plasmid stability and yield. If a plasmid is prone to recombination Use ... a strain optimized for plasmids with repeat elements, such as viral plasmids with inverted terminal repeats...been engineered to reduce plasmid recombination, but are also endA+. endA encodes a thermostable periplasmic...

...Plasmid pumpkins, team costumes, and spooky science stories. Yes, please! At Addgene, we love finding... contest trophy. We’ve had teams dressed up as plasmids, Blugene, and the Addgene ping pong table. This... 813-816. Gilbert, Lawrence I. "Halloween genes encode P450 enzymes that mediate steroid hormone biosynthesis... appearance there too) and shared fascinatingly creepy science stories. Here are some of our favorite ...

...used to create this important collection.  Creating the Plasmids To begin constructing the plasmids for ...expression plasmids were DNA sequenced to determine the VH and VL sequence. These plasmids have been deposited...scientific community with the plasmid sequences available from each plasmid webpage. Importantly, the methods...found that they could modify the CH encoding sequence within the plasmid backbone to generate R-mAbs of different...Resources on the Addgene blog:  Plasmid-Based Recombinant Antibodies  Plasmids 101: Secondary Nanobody Toolbox...Fusion PCR was performed to create an amplicon that  was cloned into a plasmid backbone for expression of...cells transfected with a plasmid to express the target antigen could be screened in a high-throughput manner...

...insect cells, they are most often expressed from a plasmid in bacteria (Ahmed et al., 2012). One challenge... the selected scFvs are inserted into a vector encoding the antibody constant domain scaffold to convert...and cheaper to generate since it’s genetically encoded.   Translational applications of scFvs: Just ...Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies for neuroscience research... rounds of screening. Created with Biorender.com.   Some phage display libraries screen antibody genes...serine with dispersed hydrophilic residues for increased solubility (Monnier et al., 2013). The linker ...expression systems. scFvs are also small enough to be screened for with in vitro display methods such as phage...

...Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent Protein Guide...accounted for in your experimental design. Find nLight plasmids here! Next steps While nLightG and nLightR greatly...Lin, D., Du, J., & Li, Y. (2019). A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo... to GRABNE, and having both colors allows for increased options for multiplexing in experiments, including...both nLightG and nLightR showing significantly increased selectivity for norepinephrine over dopamine. ...

...), 339-350 (2015). PubMed. Find the Plasmids @Addgene: Plasmids for CRISPRi are described here: http...constructs to be introduced simultaneously in a single plasmid. The functionality of the RNA scaffolds follows...differentially alter the levels of five genes that encode biosynthetic enzymes for violacein, a distinctive...http://www.addgene.org/crispr/qi/ Plasmids described in  Zalatan et al. Cell  (2015) can be found here...https://www.addgene.org/browse/article/9593/ Plasmids described in Qi et al. Cell  (2013) can be found...Programs with CRISPR RNA Scaffolds." Adam has also created a video to help scientists visualize the concepts...

...and an envelope expressing plasmid, such as our second most requested plasmid of all time, pMD2.G. Find...wide range of applications. This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB... top requested plasmid from 2016. You may notice the “V2.0” at the end of this plasmid which indicates...prior version of this plasmid. Indeed the first version is the 15th most popular plasmid of all time, but ...envelope expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini....1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...vector encodes both SpCas9 and your gRNA. It’s the former version of our sixth most popular plasmid pSpCas9...

...desired mutation into your plasmid of interest by PCR generates mutated plasmids with no methylation (there...Mutagenesis to Mutate Your Plasmids Resources on Addgene.org Check Out Our Plasmid Protocols Boost Your Molecular... for purposes other than cloning or confirming plasmids. It turns out that restriction enzymes are one...endonucleases. A given prokaryote typically has genes encoding one or a few restriction modification systems ...difficult to interpret or lead you to conclude your plasmid is not correct! Conversely, enzymes such as DpnI...methyltransferases in the PCR reaction). The template plasmid, on the other hand, should be derived from a dam...methylated adenines in any GATC sequences found in the plasmid. When the PCR products are digested with DpnI, ...