.../ARS vector (Leu2) that contains dCas9 fused to N 51025 pSLQ1661-sgMUC4-E3(F+E) Lentiviral vector that...and a short DNA motif (protospacer adjacent motif or PAM, sequence: NGG) juxtaposed to the DNA complementary...CRISPR imaging 51024 pSLQ1651-sgTelomere(F+E) Lentiviral vector with sgRNA targeting human telomeres 46913...P alindromic R epeats) pathway, as an RNA-guided DNA binding platform, to repress expression of arbitrary...anhydrotetracycline (aTc)-inducible dcas9 gene on a p15A vector with chloramphenicol resistance. The second plasmid...constitutive sgRNA expression cassette on a ColE1 vector with ampicillin resistance, wherein the N20 can...constructs 46910 pHR-SFFV-dCas9-BFP Human expression vector containing SFFV promoter, dCas9 that is fused to...

...  The RNA polymerase(s) RNA is transcribed from DNA using an RNA polymerase (RNAP). In bacteria this ... as RNAP II promoters is commonly employed in lentiviral and retroviral constructs and we will discuss...discuss these in a future post on viral vector parts. Promoter specificity Aside from choosing a promoter based... Can be induced by IPTG. Commonly found in pET vectors. Very tightly regulated by the lac operators. Good...-inducer fucose Weaker. Commonly found in pBAD vectors. Good for rapid regulation and low basal expression...Browse All Plasmids 101 Posts Learn about Mammalian Vectors Read about Reporter Gene like Luciferase and GFP...

...connectors, cDNAs, and more to find the perfect combination. The EcoFlex Kit has a wide array of DNA parts ...Modular cloning is a popular DNA assembly tool used to build single- and multi-gene constructs for a ...background MoClo is a clever way to assemble several DNA parts in a specific orientation using Type IIS restriction...kit has a focus on infrastructure with shuttle vectors, several delivery systems which can be engineered...engineered into the TUs, and even viral gene silencing vectors. The Plant Parts III kit has a focus on transformation...Fragmid system was used to assess how different lentiviral plasmid backbone elements impact viral titer ...assemblies or that compare the effects of different DNA elements on the behavior of an expression construct...

...efficiency Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector or separate...nuclease that cleaves DNA heteroduplexes, and run on an agarose gel to identify DNA fragments. PCR and restriction...components to make lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable,...Components of System Application Mammalian expression vector Cas enzyme promoter can be constitutive (CMV, EF1alpha...lines (for details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression of Cas9 and/or...separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich...CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles (for details...

...via a lentiviral vector to cell lines that either stably expressed Cas9 or that had been lentiviral infected... non-targeting guides. Toxicity is likely due to DNA damage and its subsequent repair. Additionally, the...fail to account for the effects of Cas9-induced dsDNA breaks. Are off-target cutting patterns in genome-wide...function, i.e. sites that lack open chromatin marks, DNase hypersensitivity, or are in an enhancer, transcription...control for the effects of guide expression and dsDNA breaks. Figure 2: Non-targeting vs. safe-targeting...for the inherent effects, or background noise, of dsDNA breaks caused by CRISPR. See supplemental figure...cheat sheet Read our Quick Guide to All Things Lentivirus Resources on Addgene.org Check out this guide...

...interest 85586 gRNA Cloning Vector Bbs I ver. 2 An empty sgRNA expression vector. sgRNA sequence can be cloned...Suzuki Y, Sugano S, Fujii H. DNA Res. 2017 Oct 1;24(5):537-548. doi: 10.1093/dnares/dsx023. PubMed . Identification...chromatin immunoprecipitation (iChIP) and engineered DNA-binding molecule-mediated chromatin immunoprecipitation...tagged with the recognition sequences of an exogenous DNA-binding protein such as LexA are subjected to affinity...specific genomic regions are tagged with engineered DNA-binding molecules such as TAL proteins and the CRISPR...recognizing a telomere repeat. A retroviral expression vector with the puromycin resistance gene 63591 3xFN-Tel-TAL...recognizing a telomere repeat. A retroviral expression vector with the neomycin resistance gene 64325 3xFLAG-...

...technologies. Penn Vector Core AAV Vectors Many AAV vectors from the Penn Vector Core have been transferred to Addgene...of particular Penn Vector Core catalog items. Virus-associated DNA Each viral vector service request will... Viral Vector Packaging Service Addgene produces high-quality viral vector preps...In-Stock Preps We offer commonly used AAV and lentiviral vectors as in-stock preps. These high-titer preps...and we’ll make the viral vector prep you need. Packaged on Request Viral Vector Packaging Service Information...about viral vector production at Addgene, ordering information, and more. Additional Viral Vector Resources...biosafety of viral vectors, and general biosafety guidelines. Addgene Blog: Viral Vector Posts Content from...

...intermediary DNA replication step between DNA and mRNA. DNA genome replication can involve an RNA or dsDNA intermediary...Viral Vectors 101: Viral Applications Viral Vectors 101: Viruses as Biological Tools Viral Vectors 101:...create far safer viral vectors that can be used in BSL-1 or BSL-2 labs. Viral vectors are viruses that have...increase the viral vector’s carrying capacity. In order to allow the viral vectors to reproduce when experimentally...expressed via cells cultured with the vectors. This allows the viral vectors to reproduce only in experimentally...of Viral Vectors 101 posts about each type, so be sure to check our blog, or our Viral Vectors 101 eBook...Louten, 2016 Viruses with DNA genomes can either follow central dogma (gDNA to RNA to proteins) or require...

...integrase-defective lentiviral vectors - which rely on insertion of DNA by the NHEJ pathway during DNA repair of...either introduces a ssDNA nick, a dsDNA break, or is engineered not to cleave DNA and instead activates...an experiment. For CRISPR work, this often means DNA cleavage at unanticipated sites. While off-target...genomic locus must often be proximal to a PAM site for DNA cutting to occur. The RNA sequence and PAM site confer...more. While such software typically only considers DNA sequence and doesn’t take into account chromatin ...only reduce off targets where nicking/cleaving of DNA occurs, not for deadCas activator/repressor experiments...generated near each other, this will result in a DNA double strand break (DSB), the event required for...

...information into a specific DNA site. Base Edit Base editors precisely edit DNA without promoting DSB formation...visualize genomic loci. gRNAs Empty gRNA Vectors Select an empty vector for your experiment based on a variety...Request ready-to-use viral preps of select CRISPR lentiviral plasmids. CRISPR Plasmid Kits : Find groups of...Mutated Cas nickase enzymes generate single-strand DNA breaks. Prime Edit Cas nickase enzymes fused to reverse...NEW CRISPR transposases insert large fragments of DNA. Transcriptional Regulation Activate dCas9 fused ...Targeting Type VI CRISPR systems target RNA, not DNA, across multiple systems. RNA Editing Cas13 enzymes...

...regions Joshua Mendell 212936 pGL3 Basic Vector Firefly Vector for investigating regions controlling transcription... Firefly hPGK Lentiviral expression of firefly luciferase Paul Khavari 109395 EFS-Vector Firefly EF1α ...regions using Gateway cloning. Lentiviral. Jorge Ferrer 87067 pcDNA3.1-ccdB-Nanoluc NanoLuc® Creation ...Firefly CMV Lentiviral expression of firefly luciferase Eric Campeau 18964 Luciferase-pcDNA3 Firefly CMV... Firefly EF1α Lentiviral expression of firefly luciferase Javier Alcudia 113442 pcDNA3.1 NL NanoLuc® CMV...interest are cloned into a Firefly luciferase reporter vector via Gateway cloning to test for the presence of...N-terminal Nanoluc fusions using Gateway cloning. Lentiviral. Mikko Taipale 12563 PCD FL0X Firefly Insertion...

...in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). Fabritius A, et...like APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC- and Cas9...

...Browse Tetracycline Plasmids Tet-regulated Lentiviral RNAi Vectors Curated Plasmid Collections Background ...selection TetR On Wiederschain 41393 pCW57.1 Lentiviral vector for inducible expression for Gateway cloning...options rtTA On Root 11651 pLVUT-tTR-KRAB Lentiviral vector for inducible expression of transgene or shRNA...or rtTA None On Vogelstein 25735 pSLIK-Neo Lentiviral vector for inducible expression of a miR-shRNA; selection...third-generation rtTA On Fraser 44012 pInducer20 Lentiviral vector for shRNA expression; see article for additional...Tet-On 3G On Doyon 58245 pGLTR-X-GFP Single vector lentiviral Gateway RNAi system for conditional cell line...w756-1) Lentiviral reverse tetracycline-controlled transactivator 3 (rtTA3) expression vector, CMV promoter...

...Blog: Tips for Titering Your Lentiviral Preps Lentiviral Pooled Libraries All Viral Plasmids Packaging Plasmids...researcher safety. Read our Lentiviral Guide for more information about: Lentiviral components 2nd versus 3rd...Glossary of commonly used lentiviral terms Popular Lentiviral Envelope and Packaging Plasmids One safety feature...Reiser Return to Top Popular Lentiviral Transfer Plasmids The lentiviral genome (i.e., the genetic material...generation lentiviral systems Viral production Frequently asked questions about lentivirus use Glossary...cell upon infection. When lentivirus is used for research, it is the lentiviral genome that encodes genetic...table lists lentiviral transfer plasmids that can be used with 2nd and 3rd generation lentivirus technologies...

...fragments of DNA from the helper plasmids or the cell genome can be packaged inside the vectors during production...sequence all the DNA packaged inside the viral particles (Figure 1). Briefly, packaged DNA is extracted from... common to find DNA from the packaging cell genome, bacterial genomes, cloning vectors, and helper plasmids...mixed another AAV vector with AAV-68544 to validate our method. We then extracted the DNA and processed the...used to identify DNA contaminants in AAV preparations and provide information on these DNA species with exquisite... (2015) Advanced Characterization of DNA Molecules in rAAV Vector Preparations by Single-stranded Virus...blog Check out our Viral Vectors Featured Topic Page Subscribe to Viral Vectors Blog Posts Learn How We...

...ready-to-use viral vectors from some of our most popular lentiviral and AAV vectors, as well as developing...2017, we had distributed 1,000 ready-to-use viral vector preps. Today, that number has grown to over 114,600...preps. And in 2025, we’ll be expanding our viral vectors service to include AAV Packaged on Request! “I ...research community once more. Our ready-to-use viral vectors were useful for researchers wanting high quality... plasmid request came on September 8, 2004, for pcDNA-p160MBP, while the first request for a kit came ...

...(scAAV) vector. Full-length ssAAV vector genome of both polarities are rescued from the vector plasmid...adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis." Gene therapy...Addgene.org Read Our AAV Guide Find AAV Vectors Browse All Viral Vector Resources ...designed so that the single stranded DNA folds back on itself to become dsDNA in the nucleus and is a good option... Primer for Beginners Read Our Lentivirus FAQ Learn about Viral Vector Elements Resources on Addgene....Adeno-associated virus (AAV) is a single stranded, Parvoviridae DNA virus, packaged in a non-enveloped icosahedral capsid...repeats – the sequences that flank the deliverable DNA) are removed, resulting in recombinant AAVs (rAAVs...

...Adenovirus Guide Virus Biosafety Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook Adenoviruses are ...recombined into a DNA molecule that incorporates sequences from both plasmids. This DNA molecule can then...Collection of adenovirus shuttle vectors and strains Viral... of medium-sized, non-enveloped, double-stranded DNA viruses known to cause mild to severe respiratory...that contains β-gal and GFP; contains ∼10Kb genomic DNA stuffer Vogelstein 18703 pEZYshuttle Shuttle Modified...cell types with almost 100% efficiency. Unlike lentiviruses or other retroviruses , adenoviruses do not ...

...purify specific regions of DNA, image DNA in live cells, and precisely edit DNA and RNA. The ease of generating...pooled format into lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which... two formats: as liquid DNA, or in select cases, as pre-made lentivirus . DNA libraries are shipped at...homology with the target DNA. Once the Cas9-gRNA complex binds a putative DNA target, the seed sequence...will begin to anneal to the target DNA. If the seed and target DNA sequences match, the gRNA will continue...strands of the target DNA. This results in a double-strand break (DSB) within the target DNA, located ∼3–4 nucleotides...of target DNA. Cas9n increases target specificity. Since two nickases targeting opposite DNA strands are...

...called DNA transposons. DNA transposons are commonly used as tools in the lab, so we’ll focus on DNA transposons...Transposon: The transposon is the DNA sequence that moves around. This DNA sequence encodes the proteins ...BioRender.com.   Class II TEs: DNA transposons Class II TEs are also known as DNA transposons, since they do...break, and excises the transposon from the donor DNA. A DNA footprint is left behind. When the transposon-transposase...not leave behind a DNA footprint sequence after excision. piggyBac can mobilize DNA over 100 kB in size...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...TnsB, TnsC), and 4) the donor DNA or mini-transposon that contains the DNA cargo of interest flanked by...