...of multiple DNA fragments to create TAL effector nucleases. These plasmids were designed to utilize the...-containing plasmid such as pX330. This cloning strategy not only makes it easy to create a single gRNA-expressing...Golden Gate method to create CRISPR/Cas9 constructs, assemble standard plasmids parts in different combinations...Addgene’s plasmids are used with a wide variety of restriction enzyme-based cloning methods. Each method...Entry DNA overhangs may be present in the original plasmid (Option 1) or added using PCR-based amplification...produces constructs with an attB recombination scar encoding eight amino acids, but Golden Gate assembly can...gRNA-expressing plasmid, but it can also be adapted to express multiple gRNAs. Addgene has two Golden Gate-based...

...EsCas13d. Find the Yan et al. plasmids here Find the Konermann et al. Plasmids here How do Cas13d enzymes...case, Konermann et al. targeted MAPT, the gene encoding dementia-associated tau, delivering dCasRx and...S0092-8674(18)30207-1. PubMed PMID: 29551272 Plasmids from this paper are available at Addgene. Yan...S1097-2765(18)30173-4. PubMed PMID: 29551514 Plasmids from this paper are available at Addgene. Learn... Resources on Addgene.org: Find RNA Knockdown Plasmids Check out our CRISPR Guide Read our CRISPR History...RspWYL1 increases both targeted and collateral RNA degradation by RspCas13d. RspWYL1 also increased EsCas13d...Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR repeat array near a putative...

...experiments in planarians with their recent plasmid deposit. These plasmids enable two main types of experiments... cloned into plasmids to investigate for each of these planarian species. These plasmids all have a PR-T4...insert the cDNA into the desired plasmid backbone. The Sánchez lab plasmids allow students and new labs to...appropriate plasmid and dsRNA expression is activated through the  induction of T7 polymerase encoded in the...Resources at Addgene.org Find plasmids from the Sánchez lab Find plasmids for other developmental models...transcription to create dsRNA. Students can produce DNA templates via PCR off of the same plasmid and use these...phenotype that disrupts wildtype locomotion.   Create your own plasmids to investigate other genes in planarians...

...will use plasmids and viral preps from Addgene to further their work. The Association for Creatine Deficiencies...will use Addgene’s RESOLUTE plasmid collection to perform a functional screen in mammalian cells to identify... with the Rare Genomics Institute by providing plasmid-based tools for rare disease research. Congratulations...therapeutic development,” says Iyer. Since the RESOLUTE plasmid collection consists of human and murine solute ...contraction defects. The lab also plans to use CRISPR plasmids to generate deletions of candidate genes in euploid...neurological phenotypes observed.” Using genetically-encoded biosensors and iPSC-derived cortical neurons from...research questions,” Hébert says. In the spirit of plasmid sharing and open science, Hébert plans on building...

...highlight a subset of the new plasmids 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...write about a recent plasmid deposit please sign up here.     Listen to the Hot Plasmids episode! Expanding.... Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid as well as several...further probe cellular function. The CRY2clust Plasmid can be found here. Park et al. Nat Commun. 2017...several derivatives encoding alternative promoters and/or fluorescent tags are available at Addgene. Shemesh..., the Barrick lab has deposited the pDCAF3-IS5 plasmid containing a synthetic operon that enables the ...

... diverse secretory routes in various conditions. The plasmids and lentiviral vectors encoding these proteins...amazing plasmid technologies developed by our community of depositors. With over 40,000 plasmids avaliable...amazing new plasmid tools that have come through our doors in 2015. Do you have a favorite new plasmid tool ...system plasmids are available through Addgene and are already very popular. Use these plasmids to increase...tRNA/aminoacyl-tRNA synthase pair is encoded on a single plasmid and facilitates the incorporation of ... can visualize: Cell Structure Plasmids: Cell Structure Plasmids: ER mCherry-Sec61 beta BFP-Sec61...luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have been used extensively...

...that encode site-specific recombinases, which can be used to control gene expression. See our Cre-lox ...Cre-lox Plasmids page for more information about these molecular tools. Cre AAV ID Name Promoter Fluorophore...Deisseroth 55636 pAAV-EF1a-Cre EF1a none rg* Deisseroth 121675 pAAV-EF1a-fDIO-Cre EF1a none 1, 5, 8, 9, rg...107312 AAV-hSyn-mCherry-P2A-Cre-WPRE Syn mCherry 1 Yang 107738 pAAV-hSyn-Cre-P2A-dTomato Syn dTomato 1,...Service: Recombinases Cre AAV Dre AAV Flpo AAV Light-Inducible Recombinases VCre AAV Browse the following....CMV.HI.eGFP-Cre.WPRE.SV40 CMV eGFP 1, 2, 5, 8, 9 Wilson 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a mCherry...105550 pAAV.GFAP.Cre.WPRE.hGH GFAP none 5, PHPeB Wilson 196410 AAV-GfaABC1D-Cre-4x6T GfaABC1D none 5 Carmichael...

...expression. Overview of Viral Plasmids While plasmids are typically used to encode, propagate, and manipulate...been adapted into a plasmid-based technology, such that plasmids can be used to create viruses with specific...associated with better memory, plasmids can be used to create viruses that encode that protein. Then, those...for select plasmids are now available through Addgene, learn more here . Browse Viral Plasmids by Type Visit... to find plasmids that can be used to engineer various types of viruses. Lentivirus Plasmids | Guide Lentiviruses...organism. Figure 1. Using viral plasmids to generate virus. A) Viral plasmids have been engineered such that...genomes, such that they no longer encode for more virus, but instead encode for a specific gene or other research...

...apoptosis in vivo, scientists have used genetically encoded fluorescent executioner caspase reporters. Several...these diseases and disorders. You can find the plasmids for FlipGFP and FlipCherry at Addgene. References...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...protein fusions Browse the fluorescent proteins plasmid kits ...self-assembling split GFP (GFP1-10 and GFP11) to increase fluorescence 10-fold after protease activity (... (To et al., 2016). However, the lab wanted to increase the fluorescence range to better study the spatio-temporal...β-strand of GFP, hence the name FlipGFP. FlipGFP increases fluorescence 100-fold compared to other reporters...

...clone the genes from the hybridoma into a plasmid to create a recombinant antibody.     Figure...changes in the nucleic acid sequence of the antibody-encoding genes over time as the hybridoma cells divide....Resource on Addgene.org Check out Addgene’s antibody plasmid collection Read the latest on Addgene's new antibody...of Producing Monoclonal Antibodies, 1999). This creates an immortal cell population that will continue ...hybridomas for monoclonal antibody production. Image created with BioRender.com.   The antibody-producing...Generation of Hybridomas: Permanent Cell Lines Secreting Monoclonal Antibodies. https://www.ncbi.nlm.nih.gov...

...towards successful iEP reprogramming. MettI7a1 encodes an uncharacterized putative methyltransferase. ... on Addgene.org Visit our stem cell page Find plasmids for reprogramming and differentiation Check out...individual cells. Each cell is labeled with 3-4 tags to increase the diversity of the CellTag library. The fixed...detected over a 10-week period. Their expression decreased over time but they were not silenced.    Find ...retroviral overexpression cocktail yielded a three-fold increase of fully reprogrammed cells based on single-cell...

...that’s encoded on a plasmid. A nanobody is a ~150 amino acid, single domain antibody. To create their ... your own RANbodies? Great! All of the RANbody plasmids from Yamagata et al can be found here. Can’t find...consider making you own DNA ladders with these plasmids from Penn State Need help picking the right fluorescence...Resources on Addgene.org More nanobody expression plasmids can be found here Watch this video to learn more...antibody and a ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to... RANbodies are expressed in 293T cells and are secreted from cells due to their IgGκ signal peptide sequence...them. HRP RANbody staining results in a ≥10-fold increase in signal over staining with a primary/ labeled...

...Antibodies Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene ...to bind to a previously hidden epitope (right). Created withBioRender.com.   Biological Processes  ...

...cells to retain plasmids - if a cell does not inherit plasmid due to improper plasmid partitioning, it...unstabilized plasmids were retained in less than 10% of cells while plasmids containing the plasmid addiction...selection in one plasmid, they were able to create an efficient cloning tool. The plasmid pKSCTMF contains...ensure that plasmids are passed down through generations (Figure 1). These so called “plasmid addiction ...only survive if they retain the plasmid. Since their discovery, plasmid addiction systems have been used...found that if a mini-F plasmid carried a specific 700 bp segment of DNA, the plasmid was propagated better...restriction-modification systems onto plasmids allowed cell populations to retain more of these plasmids as compared to those...

...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...anti-SARS-CoV-2 antibodies. In the library, each plasmid encodes the mutated spike protein and a barcode, and...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...Hutchinson Cancer Research Center developed a pooled plasmid library encompassing all possible amino acid mutations...head over to our CRISPR Plasmids and Resources page.  A new system uses light to deactivate photocleavable... accumulation of indels to measure time. These plasmids with different promoters driving expression of...

...Fusions Plasmids 101: Secondary Nanobody Toolbox Resources on Addgene.org View the Antibody plasmid collection...The Seeger lab found a way around this by using plasmids to generate sybodies, synthetic nanobodies against...nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target epitopes that...form of mRNA for academic research. The template plasmids for generating your own libraries are also available...available in the Sybody Generation Toolbox plasmid kit: SB_concave, SB_loop, SB_convex. These scaffolds...libraries. Each mRNA molecule in the library doesn’t encode a stop codon, which causes the ribosome to pause...efficiently subcloned into the phagemid pDX_init plasmid, using the fragment exchange (FX) cloning technique...

...for expression of a gene carried on the transfer plasmid or quantifying the number of viral copies that ...beneficial as they ensure that the correct transfer plasmid was used in the transfection; if preparing several... that comes from amplification of the original plasmid which can over-inflate results. A packing vector...number of target cells that are positive for vector encoded transgene expression following transduction with...antibiotic resistance genes carried on transfer plasmids. Target cells are transduced with serial dilutions... How Lentivirus is Used in CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used for Lineage...

...Browse our CRISPR interference plasmids Find bacterial CRISPR plasmids Read our CRISPR guide ...transposon system. This system has two parts: a plasmid expressing the Tn7 transposition genes (tnsA, tnsB... tnsB, tnsC, tnsD, and tnsE), and a plasmid that contains the Mobile CRISPRi cassette flanked by left ...copy of the RP4 transfer machinery mobilize these plasmids for transfer to recipient cells during a tri-parental...Mobile-CRISPRi ICE element forms a conjugative plasmid-like DNA circle and is transferred to recipient...transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes are transferred...RP4 transfer machinery, which mobilizes the Tn7 plasmids, and lack the ability to synthesize diaminopimelic...

...performed by transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains a site for inserting...chosen for both Cas9 and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target...targets with similar efficiency as compared to plasmid-based expression of Cas9/gRNA and can be used for... as CMV or EF1A promoters found in many CRISPR plasmids) are not expressed. Because this method does ...fluorophore tagged dCas9 may require the use of plasmid or viral-mediated delivery. You’ve decided to...and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex Schier’s Lab...experimental systems that may not be amenable to plasmid based methods. One of the most common techniques...

...the endogenous target, and thus no genetically encoded tag is required. These dyes can also be attached...® and HaloTag®. These consist of a genetically encoded enzyme variant tag that covalently reacts with ...cell and protein analysis products. You can find plasmids deposited by Promega here.    References and ...More resources from Addgene Fluorescent protein plasmids and resourcesA practical approach to choosing ...fluorescent dyes, and fluorescent ligands. Figure created with biorender.com. Fluorescence refers to...fluorescent protein tags, such as higher specificity, increased versatility in probe attachment, and compatibility...exciting the fluorophore at peak wavelength can also decrease the rate of photobleaching. Interaction with receptors...