We narrowed to 182 results for: cre encoding plasmid

... Check out All the Plasmids from the Collin's Lab Find Synthetic Biology Plasmids for Your Next Experiment...deposited the best performing sensors and constructs encoding corresponding portions of the Zika virus genome...Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from the publication at Addgene 2. Green, Alexander...Experiment Deposit Your Plasmids with Addgene ...parallel. More broadly, we drew on the combined creativity and expertise of the group and yet never hesitated...

...transcription from the accessory plasmid (AP) encoding phage gene III. The accessory plasmid library contained all...mutagenesis plasmid (MP6) permits quick, large-scale mutation of the gene of interest. They created a selection...doi: 10.1038/nature26155 PubMed PMID: 29512652 Plasmids from this paper are available at Addgene.  Additional...Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn about CRISPR History...genome-wide off-targets, since increasing PAM flexibility might also increase off-target activity. Surprisingly...gone one step further, using directed evolution to create xCas9, an enzyme recognizing a broad range of PAMs...PAMs like NG, GAA, and GAT, but also displaying increased editing specificity. We’re excited to learn more...

...Gonçalves and Ventura labs’ plasmids, including protocols, check out the plasmids at Addgene: pAdSh.PGK.Cas9...looking for a broader range of CRISPRs plasmid tools, find more plasmids, CRISPR technology guides, FAQs, and...mutagenesis similar to those achieved by isogenic AdVs encoding TALENs targeting the same chromosomal region. ...description of their experimental protocol. The three plasmids which have been deposited to Addgene are: pAdSh.PGK.Cas9...templates in somatic cells of adult animals in order to create animal models for particular cancer. That’s exactly...the Memorial Sloan Kettering Cancer Center did to create a model of Eml4–Alk-driven lung cancer (Maddalo... Addgene and could be useful to labs wishing to create this kind of animal models. Start using adenoviral...

...available Addgene plasmids he or she can use to study it (as seen in the screenshot below). Currently...refers to 3 proteins (H-Ras, K-Ras, and N-Ras) encoded by a family of proto-oncogenes. Ras proteins are...generation of mutant cell lines. Further, for each plasmid in the kit, the transcript most commonly found ...initiative to curate our collection of cancer-related plasmids such that each is linked to a known major pathway...pathway. Now, using our new Cancer Research Plasmids and Resources page, investigators may select a pathway...Visit our Cancer Collection Pages Deposit Your Plasmids for Studying Cancer Browse Our Other Collections...as 30% of all cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations...

... target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains...pCAG-EGXXFP plasmid along with the individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ...effective ones to increase your chances of gene modification in vivo. Find Plasmids for Testing Your gRNAs... system floating around, and many pre-designed plasmids containing these variants ready for purchase. ...multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and... you can insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates...

...beneficial for plasmid clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only.... pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... 13 plasmids with 14 rounds of transformation in 30 days. Figure 2: (A) MUCICAT plasmids for ... original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive...the pTargetF construct and then introduce both plasmids into the cell. The edit in the chromosome can ...the pMB1 replicon, resulting in cutting of the plasmid. At this point, subsequent introductions of pTargetF...addition of the sgRNA sequence in pEcgRNA. Next both plasmids are transformed in E. coli with the donor DNA ...

...s lab at UC Davis is to generate a genetically encoded fluorescent sensor for serotonin. According to ...Addgene.org Find fluorescent protein biosensors Browse Plasmids from the Tsien lab Browse the Michael Davidson...site-saturation mutagenesis, and medium-throughput screening, she increased the affinity of the sensor for serotonin...overwhelming response and were excited to see the many creative uses of fluorescent proteins. We’re happy to announce...thousands of protein variants simultaneously  With increasing sequencing power, scientists are able to quickly... protein-coding pathogenic human variants are decreased in protein abundance, possibly due to misfolding...neuromodulator, and evidence suggests that it is incredibly important for nearly every aspect of life.” Unger...

...Antibodies Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene ...could mean needing to increase crosslinking to counteract the effects of increased sample agitation; a ...such as qPCR or next-gen sequencing. ChIP is an incredibly useful application for exploring these interactions...crosslinking, which uses formaldehyde or UV light to create strong but reversible bonds. This will keep the... necessary for most ChIP experiences, but the increased binding can mask the epitopes antibodies recognize...shorter sonication time could be an opportunity to decrease crosslinking. (Homogenization for a tissue sample...

...al., 2007). Brainbow uses a Cre/lox system to label neurons. Genes encoding the fluorescent proteins are...fluorescent protein genes, Cre recombination is required before fluorescence can occur. Cre recombinase can be...developing fish via Cre protein microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier...gene directly following the promoter is expressed. Cre-mediated deletion of one or several of these genes...system. One such system is Brainbow 2.0, which uses Cre to trigger the inversion of two genes aligned head-to-head...head-to-head. Another variation, Brainbow 2.1, uses Cre to trigger the inversion of two tandem sets of genes...a specific tissue. XFPs are introduced using a plasmid construct called Zebrabow-GateDest containing an...

... resources on the Addgene blog Plasmid 101 Protein Expression Plasmid 101 Biotinylation Other resources...vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the development of BioID and... specific locations in the cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate peroxidase... They named this mutant APEX2.  Find the APEX2 plasmids! TurboID and minturbo: Rapid and non-toxic proximity...allowing for more precise temporal control.  Find plasmids for TurboID and miniturbo Split-BioID: A conditional...information, read the Split-BioID blog post. Find plasmids for Split-BioID RaPID: RNA-protein interaction...biotinylated. Read more about RapID in this blog! Find plasmids for RapID EXCELL: Proximity labeling for the detection...

...until now. Alice Ting’s lab has developed a new, plasmid-based biosensor to visualise changes in histone... Cindy Y. Jao, and Alice Y. Ting. "Genetically encoded fluorescent reporters of histone methylation in...Additional Resources at Addgene Find the Ting Lab plasmids at Addgene Browse More Fluorescent Protein Biosensors...grandsons that decreased their likelihood of death from cardiovascular disease but increased their risk of... DNA methylation patterns, either large increases or decreases. Hypermethylation of CpG islands in the... units of the FRET pair together and causes an increase in the FRET signal. Subsequent demethylation separates...

...efficiency of plasmid delivery to a cell. In contrast, TREE provides a read out for plasmid delivery and... histidine at amino acid 66. This histidine is encoded by a ‘CAC’ codon that is converted to a ‘TAC’ or...antibiotic resistance markers, and other enzymes such as Cre-recombinase drastically expanding the base-editing...Addgene.org Browse the CRISPR guide Find CRISPR plasmids for your research ...Base editors create specific point mutations in the genome, but they’re inefficient compared to CRISPR...experiments. In the past few years, scientists have created an array of base editing reporters that can help...Cas9-mediated editing in real time The Harris lab created ACE, a reporter that monitors APOBEC (a family ...

...and general DNA repair Reduces plasmid recombination, increases plasmid stability recBCD Exonuclease...mutation Improves plasmid yield F Host does (F') or does not (F-) contain the fertility plasmid. A low copy-number...copy-number plasmid, encodes proteins needed for bacterial conjugation. Genes listed on F´ are wild-type...Vectors Looking for More Plasmids 101 Articles?  Find them all at our Plasmids 101 Topic Page!     ...make major improvements such as those that increase plasmid yield and/or DNA quality. Table 1 below outlines...Exonuclease V activity abolished Increased plasmid stability, reduced recombination relA or relA1 Relaxed phenotype...General cloning and storage of common plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 ...

... Addgene’s experience helping scientists share plasmids, especially those of the CRISPR variety (see slides...is in one of the top 10 countries in number of plasmid requests from Addgene. Addgene - Canton Nucleic...transcriptome sequencing methods including work done by the ENCODE project consortium did not do a good job of seeing...resulted in a cell migration defect. They have created a circRNA website so scientists can view public...developments. Using the Tet-on system, his lab has created a tetracycline cas9 inducible knock-out system....

...HDR efficiency with circular plasmid templates is generally low; to increase the frequency of edits, researchers...of a homologous donor (e.g., sister chromatid, plasmid, etc.), and consists of three pathways: classical...disrupted in the donor. Choosing a type of donor: plasmid, oligo, double or single stranded? The size of ...longer than 200 bases can be difficult, thus dsDNA plasmids are generally preferred for large insertions such...researchers have designed self-cleaving plasmids that liberate the targeting region from the vector. Alternatively...these are generally more toxic to cells than their plasmid counterparts. Another consideration for dsDNA templates...., 2017). Easi-CRISPR in vitro transcribes RNA encoding your repair template, then uses reverse transcriptase...

...Antibodies Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene   ...this approach can greatly increase the sensitivity of an assay by increasing the number of signaling molecules...green; and proteins are represented in orange. Created with BioRender.com.    How do secondary antibodies...shorter than that of the primary antibody, due to increased binding affinity, many indirect antibody protocols...are designed to take place over multiple days. Increased run time is considered one of the major drawbacks...specific isotype subclasses of primary antibodies can increase the possible number of target proteins.   The ...immunohistochemistry advances have found ways to increase signals from conjugated primary antibodies. The...

...Antibodies Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene Topics...basic guidelines below, you’ll likely be able to create a small, usable panel.  First, fluorophores vary...

...realized we could apply the lessons learned from our plasmid lab to develop a reliable and robust high quality...offer many resources for producing vectors from plasmids — including the protocols we use in our lab — ...while viral vectors are not as easy to ship as plasmids, they still have a global reach. From 2016–2023...vectors in our repository are produced from only 413 plasmids containing a genetic payload. While AAV9 and AAV1...having reliable ready-to-use vectors made from plasmids in the Addgene catalog would be broadly useful...simple as it seems. Take, for instance, the GCaMP-encoding viral vectors available in the repository. GCaMP...conditions. Sharing our experiences Addgene’s plasmid repository gave us the experience to launch a high-quality...

...both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected at a favorable ratio to...Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org...Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...amount of agarose/DMEM should be proportionately increased). For some cells, add a drop of serum to aid in...up, most will eventually grow within a week. We screen the wells in groups as they become confluent, taking...important to have a functional assay in place to screen your clones. Depending on the protein, we have ...SH-SY5Y cells, as these cells did not seem to form discrete colonies but constantly migrated away from each...

... entire adult brain, using a plasmid that expresses a genetically encoded calcium sensor, GCaMP6 (Nguyen... eBook Read our plasmids 101 blog series Resources on Addgene.org Browse our plasmid collection pages...developing zebrafish pancreas. They first tagged these cells using plasmids. Then using HOTcre, a method that... resistant to chemotherapy. You can find these plasmids at Addgene. Another advantage of mice as model... feeding the worm bacteria expressing specific plasmids (L4440) that contain your gene of interest (Fire...al., 2015). If you are interested in C. elegans plasmids and resources check out Addgene’s Worm Expression...pages Read our molecular biology reference Find plasmids for your research ...