...want delivered to your target cells. This delivered DNA/RNA can then be either permanently integrated into...sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily...transfection and random genomic integration of a plasmid. Viral transduction is also generally less toxic...the virus are now delivered in other forms (mRNA, DNA, etc.) to raise immunity. Viral vectors are being...

...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...ready-to-use viral preps of select CRISPR lentiviral plasmids Additional resources on the Addgene Blog  How...

...with some plasmids. You just need three plasmids to start making AAV:  the packaging plasmid which contains... transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids have two ...of an AAV transfer plasmid. Parts shown in green are key elements of an AAV plasmid. Parts shown in grey... elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal ...transfer plasmids have a promoter, but this element isn’t required when the transfer plasmid harbors non-coding...Transfer Plasmid video!   Now you’ve got a basic understanding of some common AAV plasmid elements!...structural and packaging genes, the adenoviral helper plasmid which contains the proteins needed for the virus...

...Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria." Plasmid 51.3 (2004): 246-255. PubMed...exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate off-target mutations...a mutated allele introduced from a conjugative plasmid replaces the chromosomal copy. This is a versatile... These two steps are (Figure 1): The initial plasmid integration step (the so-called “single crossover...versatile, synthetic MCS (designed with the help of the DNA tuner program) (Latynski and Valentovich 2014). We... an allelic exchange experiment? Find the pTOX plasmids here! Continued development of new allelic exchange...PMCID: PMC4513841. Latynski US, Valentovich LN. "DNA tuner: a computer program for the construction of...

...giving you more choice in how you receive plasmids — as DNA or bacterial stabs — ‌and reinventing our ...accelerating scientific discovery. Over 150,000 plasmids empowering researchers worldwide. Countless breakthroughs...countries enriched our repository with 15,020 new plasmids — a testament to the scientific community's commitment...next-generation sequencing, setting new standards for plasmid quality control. Through our partnership with the... viral vector options. Almost any AAV transfer plasmid in our repository can now be packaged on demand...sequences and characterization data for over 150,000 plasmids, empowering bioinformaticians, data scientists...developers to unlock the potential of our extensive plasmid database — because we believe that innovation flourishes...

...components: GBPa-VP16 (activation domain); GBPb-GAL4 (DNA-binding domain); and a UAS-driven luciferase reporter...systems. T-DDOGs can also be designed with other DNA-binding domains, including the commonly used LexA...23953120. PubMed Central PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan...2015): 1334-1341. PubMed PMID: 26258682. Find plasmids from this publication at Addgene. 3. Kirchhofer...

...introducing double stranded breaks (DSBs) into genomic DNA in many cell types and often results in biallelic...homology-directed repair (HDR), a distinct cellular DNA repair pathway. Accomplishing this usually involves...involves simultaneously introducing a homologous DNA repair template, such as a sin­gle-stranded oligodeoxynucleotide...at all, such as when you are editing a non-coding DNA region? To this end we developed a technology called...Ribonucleoproteins Resources on Addgene.org Browse All CRISPR Plasmids Check Out Our CRISPR Guide Pages Find Validated...

... function (Pan et al., 2018). cDNA libraries, which consist of plasmids that over-express coding sequences...been most widely used to create specific edits in DNA, there has also been significant work on constructing... any gene by fusing mutants of Cas9 deficient in DNA cutting activity to a transcriptional activation ... activators, similar tools were made using other DNA binding proteins such as zinc fingers (ZF) and TAL... several popular CRISPR-based tools for altering DNA sequence or regulating gene expression.     References...they want to activate, we would recommend using a cDNA overexpression vector rather than going through ...and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to study in cis regulation and also...

...techniques are still used and efficiently transfer plasmid DNA efficiently into dividing immortalized cells ...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes...crossover event between the vector plasmid and the packaging plasmid caused the Psi sequence to be placed...expression of the transgene   Transfer plasmids The transfer plasmid is the vector you clone your gene of...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...reach their limits when it comes to transferring DNA into primary cells or non-dividing cells. To target... (Gag, Pol, Tat, Rev and Env) and the transfer plasmid encoding the insert of interest as well as sequences...

...incorporate new information into DNA. This also requires an exogenous DNA template. HDR, however, is a very...so-called base editor Cas9, which makes DNA changes in the absence of dsDNA breaks (Rees et al., 2018). For C...made it easier than ever both to engineer specific DNA edits and to perform functional screens to identify...efficacious to target the coding or non-coding strand of DNA. As a rule of thumb, we avoid target sites that code...CRISPR Basics with Our CRISPR Guide Find CRISPR Plasmids for Your Research Find CRISPR Pooled Libraries...technology is usually accomplished by Cas9-mediated dsDNA breaks: following a cut, the error-prone nature ...although this is rarely done). When targeting a dsDNA break for HDR, the choice of target site is far ...

...flanking the DNA-binding cleft of Cas9, the fluorescent domains dimerize and prevent DNA binding in the... cell, the dCas9-CIB1 fusion binds to the target DNA sequence as directed by the guide RNA (gRNA), while...unveiled a photoactivatable system to cleave a target DNA sequence (Nihongaki, et al., 2015) resulting in a... Additionally, high intensity exposure can cause DNA damage. Far-red light, however, can penetrate 5 mm...fluorescent reporter gene in mice using Minicircle DNA vectors encoding the FAST constructs. This proof-...stimulation, the pdDronpa moieties move apart, allowing DNA binding and cleavage. This technique was applied ...deactivated the protein’s ability to cleave targeted DNA, settling on photocaging the K866 lysine, as seen...

...will be able to request an AAV composed of an AAV plasmid in the Addgene repository and their choice of five... more.  What will be available?  Over 3000 AAV plasmids in the Addgene repository will be available for...Packaged on Request service. A small subset of AAV plasmids will have technical or legal restrictions that...both our depositors and requestors.  Requesting Plasmids available for Packaged on Request consideration...sequencing to confirm the identity of the packaged DNA and serotype. Please note that Addgene cannot perform...recommend you check the original paper of your AAV plasmid of choice for more information on its performance...performance.   Titers We expect titers for plasmids packaged in AAV1, AAV5, AAV8, and AAV Retrograde to be 1x1013...

...Addgene-Seeding Labs plasmid grant is a collaborative endeavor providing plasmids to researchers in developing...Addgene-Seeding Labs Plasmid Grant Check Out Previous Addgene-Seeding Labs Plasmid Grant Awardees Listen...scientists who were awarded the Addgene-Seeding Labs Plasmid grant this year. These scientists work on projects...technology to engineer plants that are able to destroy DNA viruses, similar to the anti-phage CRISPR effect ...as the pET GFP LIC cloning vector and the pOKD plasmids. The recombinant Cry proteins will then be screened...HyPer3 (2) which he will co-express together with a plasmid encoding glycine oxidase. The goal is to determine...excited to see how all of the Addgene-Seeding Labs Plasmid Grant awardees' projects progress. Subscribe to...

.... pBLADEONLY: This plasmid contains only BLADE and is based on the pTRC99a plasmid. It does not include...AraC and other promoter systems, head over to our Plasmids 101: Inducible Promoters article.) Modifying AraC...team tested different linkers between VVD and the DNA-binding domain of AraC using a mCherry reporter downstream...reporter downstream of the PBAD promoter. To use this plasmid with your gene of interest, you would replace mCherry...PBAD promoter. If you already have a pBAD33-based plasmid expressing your gene of interest, all you have ... and shared with Addgene. You can find all the plasmids deposited from Di Ventura's lab, including the...nuclear export system (LEXY) here. Optogenetic plasmids from the Khammash lab can also be found here. ...

...., 2019). AdVs are non-enveloped double-stranded DNA viruses, with a linear genome between 26–45 kb in...initiating the whole process – such as the viral DNA polymerase – it makes sense that vectorization strategies...strategies include creating an AdV transfer plasmid with the E1 and E3 (Early 1 and Early 3) genes deleted.... These genes’ functions are rescued by helper plasmids used during production and/or by genes present...made available by this deletion from the genomic plasmid can be used to house a recombinant sequence (transgene...late genes are usually present in the transfer plasmid for the majority of adenoviral vector systems used...

...Browse fluorescent protein plasmids and resources See all Jaffrey lab plasmids ...winter day, you should be happy to learn that many DNA-based reagents are named for soup ingredients! From... cellular processes by transforming cells with plasmids encoding aptamer-tagged RNA and later introducing...aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment...has been used to investigate intracellular viral DNA dynamics in neuronal cells (Nilaratanakul et al.,...

... that a small DNA binding motif in MECP2 interacts with the minor groove of AT-rich DNA sequences to regulate...their MECP2-related plasmid tools with Addgene and share additional resources. Plasmid tools to facilitate...to partner with them to launch a Rett Syndrome Plasmid Collection and Resource Center for Rett Syndrome... Rett Syndrome in novel ways.  Addgene’s MECP2 Plasmid Collection and Resource Center will continue to...

...multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the...rectangles) are cloned into various plasmids using oligonucleotides. These plasmids contain Type IIS restriction...different purposes.   Gersbach Lab multiplexing plasmids: This plasmid set allows you to express 2-4 gRNAs, with...assembled to produce a plasmid encoding the gRNAs and Cas9. As with the Gersbach lab plasmids, multiple Cas9 ...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids can be used to express up to 8 gRNAs after...Yang Lab Single Transcript Multiplexing Plasmids: These plasmids are similar to the Csy4 polycistronic ...

...s incredible.” Looking for CRISPR Plasmids? CRISPR-Cas9 Plasmids available at Addgene CRISPR-Cas9 References...Forbes respectively: "A Powerful New Way to Edit DNA" and "This Protein Could Change Biotech Forever."...Last month, the Lawrence Berkeley Lab's Jennifer Doudna and Eva Nogales presented the first detailed picture...

...the edited strand and wild-type strand of the host DNA. When these strands hybridize, the nuclease can detect...account other elements, including those relating to DNA structure (e.g. epigenetic modification, bulges) ... et al. "High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity...Frock, Richard L., et al. "Genome-wide detection of DNA double-stranded breaks induced by engineered nucleases... Crosetto, Nicola, et al. "Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing...Addgene.org View Our CRISPR Guide Pages Find CRISPR Plasmids for Your Experiments Get Ready-to-Use CRISPR Lentivirus...