...Chayama K, Yamamoto T. Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400. PMID: 24954249 Find the Mulitplex...University who described their new tool in Scientific Reports in June. “The multiplexity is one of the most advantageous...will also allow simultaneous activation and/or repression of multiple genes, and more. The researchers ...

...Mapping Multiprotein Complexes in Human Cells. Cell Rep . 13(3):621-33. PMID: 26456817 Fu Y, Sander JD, Reyon...other hand, if a repair template with homology arms was supplied, the break could be repaired according to...loci. Activate or Repress : dCas9 fused to transcriptional activators or repressors can modulate transcription...mouse, and rat) Bacteria ( E. coli , Streptococcus , Streptomyces , and others) Drosophila Plants (monocots...Clustered Regularly Interspaced Short Palindromic Repeat) sequences were initially discovered in the E. ...Protospacers are separated by short palindromic repeat sequences. (1) The CRISPR array is transcribed ...tracrRNA) with homology to the short palindromic repeat. The tracrRNA helps recruit RNAse III and Cas9 ...

...Staphylococcus aureus Cas9. Sci Rep 6: . https://doi.org/10.1038/srep32565 Resources on the Addgene Blog...Blastocyst and CRISPR/Zygote Injection Approaches. Sci Rep 8: . https://doi.org/10.1038/s41598-018-33408-9 Mizuno...system derived from Streptococcus pyogenes. An alternative Cas9, from Streptococcus aureus, is as effective...out an “SOS” signal and cellular repair mechanisms rush in to repair the damage. If all they can do is...homology directed repair pathway (HDR), be it a single nucleotide change, insertion of a reporter gene, or replacement...confused. It was supposed to be hard. I was mentally prepared to go through a learning curve that would result... been following the Addgene blog, it has been repurposed for genome editing in eukaryotes, with the most...

...proteins. Krumholz et al (Sci Rep. 2014 Feb 3; 4:3939. doi: 10.1038/srep03939.)  ...background.” Just last month in Scientific Reports, Verkhusha’s team reported another advance in the application.... Transparency window That’s exactly what they reported doing in a paper that appeared in Nature Biotechnology...

...mediated gene disruption in the HIV provirus. Sci. Rep. 6, 20064 (2016). PubMed PMID: 26829887. PubMed Central...donor sequence integrated through homology-directed repair (HDR) (1), or indel mutations resulting from non-homologous...detection limits of about 5% mutant targets have been reported for mismatch nuclease assays (2, 6, 7). Ability...

...immunoglobulin binding domains.  Santilli, R.T., et al. Sci Rep (2021). https://doi.org/10.1038/s41598-021-96051-...a subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These...FLAG is eight amino acids, HA is nine), multiple repeats of the tag can recruit multiple frankenbodies to...induce high levels of gene activation for both reporter and endogenous genes. It is highly specific in...

...Gene Expression Analysis During iPS Reprogramming Process. Sci Rep 8, 8716 (2018). https://doi.org/10.1038...variable nonsense to publication quality data. Sci Rep 7, 2409 (2017). https://doi.org/10.1038/s41598-017...ensure you’re recording and reporting the right information so others can reproduce your efforts. And if you...step. Scorpion chemistry in particular has been reported to work well in the Lightcycler while being relatively...Yes Agarose gel Sunrise primers   Low Reporter and quencher on hairpin structure included in ...much as sixfold variation in expression levels reported (Biassoni, 2014), so you’ll want to ensure that...steady pair of hands to produce reliable and reproducible data. Here are some tips to help you in your...

...bfloGFPc1. Bomati et al., Sci Rep 2014 Jun 27;4:5469. MitoTimer plasmids to report mitochondrial turnover ...the cells will turn green. Mashiko et al., Sci Rep 2013 Nov 27; 3:3355. Rinehart lab reagents for...protocol for TALEN construction. Sakuma et al., Sci Rep. 2013. Nov 29;3:3379. Interested in more genome...and 13 in the TALE repeat, other naturally-occurring variations in the TALE repeat, referred to as “non-RVD...an identical variable repeat (VR) at the 4th and 32nd residues of the TALE repeat and the VR differs between...mutagenesis libraries or to insert heterologous genes, reporters, or other markers into a target genome. They did...Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer) to...

...for multiplexed super-resolution microscopy. Cell Rep Methods 2(10):100301. https://doi.org/10.1016/j.crmeth...by reducing the distance between the target and reporter. They have fewer non-specific binding opportunities...it either needs to be directly conjugated to a reporter or you need a light-chain specific conjugated ...fairly easy to find these in a Fab fragment format. Reporter-conjugated anti-IgG Fab fragments make good secondaries...

...question may take some time. If the customer service rep does not tell you upfront how long it will take to...

...Monoclonal Antibody Sequences. Sci Rep 6: . https://doi.org/10.1038/srep31730 Tsuruta LR, dos ML, Moro AM (...recombinant antibodies are essential for good reproducibility. Non-recombinant antibodies are notorious for... for being the source of irreproducible data (Begley & Ellis, 2012; Baker, 2015). This contributes to ...doi.org/10.7554/elife.43322 Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature ...10.1038/483531a Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature...

...for high-titer lentivirus preparations without ultracentrifugation. Sci Rep. 2015; 5:13875. PubMed PMID...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from... addition, the promoter of the 5’ long terminal repeat (LTR) is mutated in third generation systems such...considerably lower volume than with the unconcentrated prep. For example, at Addgene we typically concentrate...concentrate 100-fold meaning that a 10 cm2 dish preparation of virus will only yield 150 μl of material as opposed...opposed to the 15 ml collected from an unconcentrated prep. With this in mind, if the experiment requires ...

... the SAM system differ in target preference. Sci Rep 9: . https://doi.org/10.1038/s41598-019-54179-x Tanenbaum...single copy of VP64 per each dCas9, SunTag uses a repeating peptide array to fused with multiple copies of...A, Zhang F, Marraffini LA (2013) Programmable repression and activation of bacterial gene expression using...Doudna JA, Weissman JS, Arkin AP, Lim WA (2013) Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific...

...Degron Tagging with Short Homology Donors.” Cell Rep.15(1) (2016):210-8. PubMed PMID: 27052166. Kunzelmann...including protein enhancers and repressors, can be included in the repair template for inducible alteration...overexpressed fluorescent protein fusions is to replace the genomic copy of your gene of interest with ...site and, using the endogenous homology directed repair pathway, a new user-defined DNA sequence (like ...to individually generate gRNAs and homology arm repair templates containing the tag for each insertion...sequence, as well as the 5’ and 3’ homology arm repair template (Fig. 1, Step 1). There is no need for...expression and the second site accepts the homology arm repair template. SapTrap includes a prebuilt donor plasmid...

...Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids from...methods for repairing DNA after a double-stranded break. HDR copies the sequence from a repair template ...HDR, and MMEJ repair varies from organism to organism. MMEJ doesn’t yield a “perfect repair” like HDR, but...end-joining (NHEJ) to make deletions or homology-directed repair (HDR) to create precise genome edits. But have ...have you heard of another double-stranded break repair mechanism: MMEJ (microhomology-mediated end-joining... only very small homology regions (5-25 bp) for repair, making it easier to construct targeting vectors...with flanking sequence homology for error-free DSB repair. NHEJ joins the ends of a DSB in an error-prone...

...bp, plus a 20 bp barcode 400 ng Solid pCMV-Rep 78/68 SSM Library Amplification and Sequencing Protocol...amplification protocol with your deposit. This is because repeated amplifications of pooled libraries can lead to...containing smaller, recombined backbones that may replicate faster. For this reason, if your library has inserts...recommend ensuring the cells used are meant for repetitive sequences (such as STBL4 or Endura Duo), growing...Protoc. 2017 with modifications Screening pCMV-Rep78/68 Scanning Saturation Mutagenesis (SSM) Pooled ...Addgene blog Genome-Wide Screening Using CRISPR Preparing Pooled Libraries NGS Quality Control for Pooled...

...fluorescent voltage sensor to detect neural activity. Sci Rep 9, 15878 (2019). doi: 10.1038/s41598-019-52370-8 ...sensors are better classified by their fluorescent reporter and the mechanism of their voltage sensing component...

...diverse array of human cells. Sci Rep 4: . https://doi.org/10.1038/srep05105 Stephens CJ, Lauron EJ, Kashentseva...Scientific Reports paper introducing the delivery method in 2014, Manuel Goncalves lab report that AdV-...provide a DNA template to repair the double strand break via homology directed repair and you would probably...years for in vivo delivery and gene therapy and represent one of the most studied viruses. Adenoviruses ...utilized the Non Homologous End Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an ...approach to NHEJ is to utilize the Homology Directed Repair (HDR) to knock in corrective genes. For this you...for any such disease. This technology therefore represents a potential platform that could be realized via...

...Gomez-Nicola D, Riecken K, Fehse B, Perry VH. Sci Rep. 2014 Dec 22;4:7520. PubMed PMID: 25531807. PubMed...desired transgene flanked by HIV-1 long terminal repeats (LTRs) that facilitate viral packaging and host...transduced rat neurons in vivo with beta-gal and other reporters, and they showed that expression persisted for...original system, the possibility of creating a replication-competent virus via recombination between the...of recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through...

...performed using the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding...through how we produce both our AAV preps and our lentiviral preps.   Viruses are generated using standard...order to generate high quality preparations. After production, all virus preps are titered and subjected to...-containing) AAV particles within representative vector preparations was quantified with electron microscopy... lentiviral preparations (“-LVC” catalog items) are generated from lentiviral preparations (described ... to the cleared viral preparation pool. In addition, the final viral preparation undergoes PCR with primers...Addgene has a growing catalogue of viral preps available to request. In this post, we’ll walk you through...