...) ChiRP-seq RNA Hi-C DNA (genome-to-genome interactions) DNase-seq DNA (regions that are accessible... regions of DNA that demonstrate “spooky action at a distance.” Through the marvel of DNA compaction, ...details on DNA compaction and the structural proteins that make it possible, see the appendix DNA is a 3D...millions of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus in the same family...See below for further details. DNA is a 3D Fractal Figure 2: DNA compaction. Graphics courtesy ...For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning of the third chromosome, ten...coordinates for genetic elements like protein, RNA, and DNA (Table 1). As with GPS in the real world, coordinates...

...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...

...macromolecules used in scientific research such as DNA or protein. It comes as no surprise then that stability...Base modifications What is more stable than RNA? DNA! One way to make your gRNA more efficient on a budget...ribonucleotides for deoxynucleotides. A partially DNA gRNA is surprisingly well-tolerated by the Cas9 protein....org  View Addgene's collection of empty gRNA vectors View Addgene's CRISPR resources Request ready-to-use...-to-use viral preps of select CRISPR lentiviral plasmids Additional resources on the Addgene Blog  How...

...  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...

...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...

...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...

...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...

...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...

...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...

...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...