...Argonaute proteins play an important role in RNA interference, binding RNA guides to cleave foreign RNA. Argonaute...choosing a genomic target. NgAgo also appears to outperform Cas9 in GC-rich regions. DNA guides NgAgo uses... 2. Swarts, Daan C., et al.  “DNA-guided DNA interference by a prokaryotic Argonaute.” Nature. 507(7491...

...that can be added when ordering them or can be performed enzymatically later. Experimental Procedure Digest...

...Addgene uses electrocompetent Stbl4 cells when performing amplifications, and we use electroporation to... distribution, with an AUC of 0.5 representing perfect uniformity (see Figure 3A). Addgene finds that ...pooled plasmid libraries. Pooled libraries can be a powerful research tool, but their proper use requires careful...

...but it doesn’t really matter if those overhangs perfectly match up. They just need enough complementary ... To overcome this limitation, one option is to perform a hierarchical assembly, assembling fragments in...Choosing a Molecular Cloning Technique Choosing Your Perfect Plasmid Backbone ...

...lab team manually transforms, innoculates, and performs DNA minipreps for small batches of samples. As...standard pipeline. For the first ~50 plasmids we perform our standard full plasmid sequencing by NGS. Addgene...and make them available to scientists without performing the full suite of time consuming, costly, and...

...Listen to the C. elegans RNAi podcast segment RNA interference (RNAi) is extensively used in C. elegans research...endogenous RNAi pathways. These mutations however can interfere with other genetic pathways making it difficult...experiments. Potential experiments that one might perform using the purified domains include protein stability...

...couple of promising variants to check how they perform under your experimental conditions. This is time...combinations of biology and equipment. Because the performance of a fluorescent protein strongly depends on ...endogenous tagged genes) can be used to verify performance under different imaging conditions (e.g. different...

...have taken the CRISPR world by storm. Like RNA interference, these enzymes can knock down RNA without altering...nuclease domains characteristic of the Cas13 superfamily. Yan et al. proceeded to study orthologs from...multiple RNA regulating methods: small hairpin RNA interference, dCas9-mediated transcriptional inhibition (...

...-based method for screens. Cas12a’s ability to perform multiplex editing within a single expression cassette...used to create a genome-wide library capable of performing similarly to Cas9 screens in a compacted library... via a nuclease-active CasMINI. This small yet powerful system opens up new possibilities in both cell...

...target DNA. Conventional thermal cycling is then performed to amplify the target using PCR. Instead of tracking...mutation status. The drop-off probe is designed to perfectly complement the wild type sequence and is not capable...design such assays, in addition to testing their performance (2). Remember that although digital PCR is a ...

...separable from that of EGFP. This makes mCyRFP1 a powerful tool for use as a donor in FRET (fluorescence ...residues on the outer surface to break dimerization interfaces (affecting many of the RFPs developed from natural...All three mScarlet variants have shown great performance in cellular functional imaging and in protein...of essential genes in vivo. Utilizing CRISPR interference (CRISPRi) technology, the arrayed knockdown ...

...of AAV could facilitate the development of more powerful AAV serotypes, and perhaps lead to controlled ...neuronal networks, improved AAV vectors could also be powerful tools for the field of gene therapy if they can..., Phillips, J.W., Hohmann, J.G., Wohnoutka, P., Gerfen, C.R., Koch, C., Bernard, A., Dang, C., Jones, ...Michael, S., Kuleshova, E., Ojala, D., Huang, C.C., Gerfen, C.R., Schiller, J., Dudman, J.T., Hantman, A.W...

... forces scientists to use invasive measures to perform in vivo optogenetic experiments. Intracranial injections...correlated with the model’s predictions.  Three top performers, named ChRger1, ChRger2, and ChRger3, were then...minimally-invasive optogenetics While ChRgers performed well in a tissue culture dish, that doesn’t guarantee...

...step to remove the tag in order to prevent it interfering with protein function. In contrast, the polyhistidine... it is, and the greater the risk that it will interfere with protein function.  Additional Benefits of...this one developed by Dr. James Trimmer’s lab, to perform assays that they otherwise would not be able to...

...stranded? Well, that’s just for DNA – right? RNA performs almost all of its biological functions in our ...sense dsRNA and respond accordingly. While not perfect, these sensing systems work to distinguish between...blog CRISPR 101: RNA Editing with Cas13  RNA Interference in Plant Biology  ...

... from Addgene’s repository that can be used to perform screens in human cells. General information regarding...Liu at Oxford University, we are now providing a powerful CRISPR-based screening source for the Drosophila...resources for fly labs who wish to harness this powerful genome engineering tool. Bassett, et al. J Genet...locus causes that locus to be methylated. This powerful tool has been used to direct methylation to and... that high-throughput tagging projects can be performed. Schwartz, et al. Genetics. 2016. PubMed PMID:...

...cloning strategy designed by synthetic biologists to perform large scale, one-tube cloning reactions. In this...courtesy of Rob Hurt. What makes MoClo systems so powerful is their modularity — many parts can be assembled...used to assemble multigene constructs capable of performing production and subsequent necessary modification...

...Andrews et al., 2019). Finally, scientists can perform the entire process of antibody selection and maturation...recombinant antibodies. The selection process is often performed via phage display, yeast display, ribosome display...they are using and any validation experiments performed will be useful in perpetuity.  Second, plasmids...

...specific to lasers or optogenetics so cheaper, less powerful alternatives to the Master-9 that only control...published first. Because behavioral optogenetics is performed in freely moving animals, the fiber optic patch...into some of the important considerations when performing optogenetics behavioral experiments. Thank you...

...thousands of designs in parallel. To do this, we performed high-throughput ‘self-targeting’ screens with ...PRIDICT2.0 score. For the best prime editing performance, we suggest that users select some of the best...learning–based model with high generalization performance. Science Advances, 5(11), eaax9249. https://doi.org...