...system consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility... First, the packaging system is split into two plasmids: one encoding Rev and one encoding Gag and Pol...transgene-expressing or knockout cell lines. Many CRISPR plasmids are designed for lentiviral use, including Addgene...impact and safety. Many of Addgene’s most requested plasmids are used with lentiviral expression systems. If...

...collection of plasmids each carrying a single gene with a defined mutation or gene fusion. Plasmids carrying...Resources on Addgene.org Find plasmids for your research Find cancer research plasmids and resources ...mutations, and the group has deposited several hundred plasmids from this study with Addgene. Read on to learn...learn more about their work, and how these plasmids can help cancer researchers studying an assortment of...cancer research community, but what else are these plasmids useful for? They could be used to more thoroughly...phenotypic effect in a different system. These plasmids can also help scientists understand how a given...

... usage bias in Plasmids 101: Codon usage bias Read more about plasmids in our Plasmids 101 blog series...There are 64 different codons that encode 20 amino acids and three stop codons, meaning that the same amino...prefer certain codons over others for certain amino acids. This is termed codon usage bias. In fact, some ...transfer RNAs (tRNAs). tRNAs bind to and deliver amino acids to the ribosome where they are incorporated into...depicts the 64 codons that encodes the 20 amino acids and three stop codons. (B) Example of codon usage...sequence databases: status for the year 2000. Nucleic Acids Research 28:292–292 . https://doi.org/10.1093/nar...usage bias, and its potential applications. Nucl Acids Res 15:1281–1295 . https://doi.org/10.1093/nar/15.3.1281...

...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...series, we’ll highlight a few of the new CRISPR plasmids available at Addgene. We will still periodically... Bioengineering Find the optogenetic repressor plasmids Cytosine base editors with minimized off-target...paper in Nature Biotechnology Find the base editor plasmids CRISPRi in Caulobacter crescentus Figure...once. Read the paper in mBio Find the CRISPRi plasmids A multiplexed gRNA piggyBac transposon system... Read the paper in Scientific Reports Find the plasmids Enhanced CRISPR-based DNA demethylation ...paper in Nature Communications Find Casilio-ME plasmids If you have a new CRISPR tool you’ve recently...

...Model Organisms blog post Read more about plasmids in our Plasmids 101 blog series Learn more about CRISPR...collection page Find more plasmids and resources for plant resources on Plant Plasmids Collection Page Read...in plasmid manipulation and genetics research. Plasmids are ideal tools for studying genetic modification...disease models.  Check out Addgene’s collection of plasmids containing Rattus norvegicus genes! Mexican axolotl...several vectors for use in the axolotl including plasmids for use in CRISPR and Tol2 transposase induced... and after embryogenesis. Addgene has over 600 plasmids that contain Xenopus genes and a tool-kit from...Read our molecular biology reference Find plasmids for your research ...

...for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and the...plant-specific modifications to commonly used CRISPR plasmids are necessary to use the CRISPR system in plant...et al., 2013). Addgene carries Cas9-containing plasmids for knockout, activation and repression of target...systems. Luckily, you don’t have to look far for plasmids that have the plant-specific modifications required...targeting your favorite plant gene; you can find many plasmids for a wide variety of CRISPR applications in plants... available through Addgene. In addition to the plasmids described above, Addgene carries several useful...Plant Parts Kit from the Patron Lab.  As with all plasmids in the repository, we highly recommend reading...

...Michael Davidson fluorescent protein collection Check out Addgene’s fluorescent protein plasmids and resources...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...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. "...

...vectors (LTRs and ITRs), plasmids with multiple polyA sites, and other plasmids that contain repeating ...I help troubleshoot involves viral vectors and plasmids with repeating sequences. Repeating sequences ...other very much undergo recombination, two smaller plasmids are formed. Viral vectors are prone to recombination...and 3’ LTR regions, it splits into two smaller plasmids. The ~3.7 kb recombined vector backbone contains...elements. Knowing how to amplify recombination-prone plasmids and testing your prep can save you the headache...browse the table of common gene mutations in our Plasmids 101: Common Lab E. coli Strains blog post. Check...copy number, try following protocols for low copy plasmids. For example, using a longer growth period (up...

...to provide scientific resources, equipment, and plasmids to researchers in resource-limited settings. More...the Addgene-Seeding Labs plasmid grant provide plasmids to these researchers. In the third round of the...biofilms formed on various structured surfaces. Plasmids expressing fluorescent proteins will be used to...microscopy and FACS. In an additional approach, plasmids such as pACBSR-hyg and pFLP-hyg will be used to...regulation and diabetes will be expressed from plasmids (find more information on bacterial expression...information processing during this condition is crucial. Plasmids expressing fluorescent proteins and a new two-...create this bacterial sensor, he will construct plasmids containing promoter regions that activate fluorescent...

...which non-canonical amino acids (ncAAs, also sometimes called unnatural amino acids) are placed in a protein...probably quite interested in the canonical 20 amino acids. But in your quest to learn more about your protein...protein of interest, you may find the available amino acids a bit…limiting. It may be time, then, to look towards...and synthetase introduced to the bacteria using plasmids. Rapid expansion occurred, with hundreds of new...Blue Flame award to identify highly requested GCE plasmids. But they’d also love scientist interested in,...and resources, and then checking out all of the plasmids and bacterial strains (available at Addgene for... Site-specific Incorporation of Unnatural Amino Acids into Proteins. Science, 244(4901), 182–188. https...

...stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, ...Researchers express genes of interest from plasmids in order to study gene function or to engineer cells...molecular biologist to do? Let’s take a look at how plasmids recently deposited by Chris Voigt’s lab help keep...gene expression and plasmid number. In a suite of plasmids modified from pSC101 with copy numbers ranging...Because pSC101 is not representative of all types of plasmids, the stabilized promoters were also tested in ..., R6K, and ColE2 plasmids) as cell division slows (Kittleson et al., 2011). Segall-Shapiro et al. showed...collections page Download the Plasmid 101 eBook See all plasmids deposited by the Voigt lab ...

...suffice. This can result in DNA:RNA hybrids. Unlike dsRNA, DNA:RNA hybrids are abundant in normal cells and...when using these reagents. RNA:DNA hybrids Persistent RNA:DNA hybrids that arise during transcription are...Endogenous sources of duplexed RNA DNA:RNA hybrids Sometimes ssRNA just needs a buddy, and even DNA...throughout a single cell, so these nucleic acid hybrids are always around. R-loops are particularly enriched...substrates and in distinct cell cycles. When RNA:DNA hybrids levels are allowed to reach very high or low levels...al., Human proteins that interact with RNA/DNA hybrids. Genome Res., 28(9): 1405-1414 (2018). 10.1101/...

...construction of new plasmids. Dr. Duedu will also use polymerases produced using plasmids from Addgene to ...doctors Bengyella and Duedu and how they plan to use plasmids from Addgene to advance their research. Louis ...cellulosic biomass. Dr. Duedu would like to use the plasmids he acquires through Addgene to produce molecular...fluorescent proteins. Dr. Duedu’s plans for these plasmids have a heavy focus on teaching. He hopes to teach...in environmental samples via PCR. We hope these plasmids will help Dr. Duedu inspire new Ghanaian scientists...and creative ideas applicants will propose for plasmids from the Addgene community and hope we can do ...

...separate (and thus smaller) plasmids. Finally, having four separate plasmids allows researchers to partially... four viral genes are each encoded on separate plasmids and are co-transfected into cells alongside a ... the same plasmid (as opposed to four separate plasmids), there are a few reasons arguing against this...Technical tips It is important that all of these plasmids (those containing NP, VP35, VP30, L, and the minigenome...extremely important. For example, particular ratios of plasmids encoding NP and VP35 allow for efficient polymerase... It is important to note that this system uses plasmids where the genes (either viral genes or minigenome...Resources on Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV ...

...enChIP Plasmids? All Hodaka Fujii lab plasmids Sign up to receive email alerts when new plasmids from ... shopping for a wide range of plasmids. The merit of depositing plasmids to Addgene has been spectacular... researchers all over the world have requested plasmids for enChIP through Addgene. Retroviral Expression...user-friendly and precise. We will deposit the plasmids of these systems to Addgene in the near future...Community Addgene: In 2012 your lab first deposited plasmids with Addgene. In 2014, you joined Addgene's Advisory...

...GFP is a ~27 kDa protein consisting of 238 amino acids derived from the crystal jellyfish Aequorea victoria...from a maturation reaction of three specific amino acids at the center of the protein (Ser65, Tyr66, and .... Scientists can easily utilize GFP-containing plasmids as a means to many functional ends. We’ve listed...each protein or domain being tested. Find FRET plasmids here. Split EGFP: An alternative to FRET, split... marking/selection: Expression constructs like plasmids often include GFP as a marker to help identify...serve as an alternative to antibiotic selection. Plasmids of this type may have the GFP under the control...techniques in fluorescent microscopy. Read More: All Plasmids 101 Articles Addgene's Fluorescent Protein Guide...

...amino acids in size, which includes SpCas9, and shortern orthologs approximately 1000 amino acids in size... Staphylococcus aureus Cas9 (SaCas9, 1053 amino acids) displayed cleavage activity in mammalian cells....small enough to package into AAV. At 984 amino acids in length, Cas9 from Campylobacter jejuni (CjCas9...Neisseria meningitidis Cas9 (NmeCas9) at 1,082 amino acids, can also be packaged into AAV. NmeCas9 has the ... proteins with AAV, at just 1,108 and 986 amino acids, respectively, the size of both is within AAV’s ...capabilities of full-length SpCas9. This set of plasmids includes AAV-Cas9C-VPR for targeted gene activation...Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids ...

...anyone who has worked with individual plasmids knows, some plasmids amplify well and others don’t. The size...pooled library, which contains a collection of plasmids in a single tube, or a pre-made viral prep library... prepare the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral...library and appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus...pooled library has hundreds to millions of distinct plasmids, all of which need to be represented in the same... verify via NGS now than to learn some of your plasmids didn’t amplify after your screen is completed!...need to infect many more cells than the number of plasmids in the library.  Spinfection If you have a large...

...Read the paper in Nucleic Acids Research Find the drug inducible Cas9 plasmids E. coli genome-wide inhibition...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...series, we’ll highlight a few of the new CRISPR plasmids available at Addgene. We will still periodically...domain needed for DNA unwinding. They’ve deposited plasmids for active and inactive CasX for E. coli and human...expression. Read the paper in Nature Find the CasX plasmids Drug inducible transcriptional activation and...

...CRIPSR Resources on Addgene.org Find ACVR1 Plasmids Find Plasmids for Disease Research on OurSpecial Collections...decided it was time to get in shape. After three kids, I had put on typical middle age weight and “exercise...Boston Marathon, he wanted to run and play with my kids like any other youngster. His parents, Kristi and...not be able to run around and play like the other kids his age. A potential FOP cure But, there is HOPE...two ways. First, Addgene is distributing ACVR1 Plasmids from a variety of research labs and hopes that...pathways that impinge upon FOP to deposit their plasmids with Addgene and join us in the fight against ...