...or disruption of normal protein function, and ultimately misinterpretation of the protein’s cellular role...high-throughput genetic screen, it is expensive and time consuming to individually generate gRNAs and homology...the Addgene Blog Learn about Fluorescent Protein Timers Check out Photoactivatable Fluorescent Proteins...

...cellular proliferation. We have appreciated for some time the causes of this uncontrolled proliferation, in...encouraging cells to grow and divide only at the right times and in the right places. Checks and balances: protecting...undergo programmed cell death while simultaneously stimulating growth and division. The combined effect is unregulated...

...developed into lentiviral vectors. Lentiviruses are sometimes referred to as “complex retroviruses ” (Cullen...cells as the virus. Because it is made at the same time as the other components of the vector particle, ...viruses and transduction by vectors (which we sometimes still call “infection” as a shorthand, although...

...foreign DNA, and the Cas9-gRNA RNP is degraded over time, using RNPs may limit the potential for off-target... Cas9-gRNA RNP will be degraded by the cell over time. As a result, RNP mediated CRISPR experiments may...B., et al. "A thermostable Cas9 with increased lifetime in human plasma." Nature Communications8.1 (2017...

... increasing HDR efficiency to read more about optimizing this distance. Homology arms and types of donors...apoptosis in as little as 10 hours, so read up on the timing of this approach carefully (Lin et al., eLife)!...homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. eLIFE. (2014) 10.7554/...

...colony PCR can accomplish the same thing in less time and for less money. The key steps to colony PCR ... on a gel   Now that your PCR is complete, it’s time to run the products on an agarose gel to determine... biology tool kit. Consider giving it a try next time you’re screening for positive clones! Tips and ...

...advantage over conventional fluorescent dyes of the time was the fact that it was non-toxic and could be ... internal ribosome entry site (IRES). This is oftentimes used in conjuction with FACS (see below). Fluorescence-activated...protein to use in your upcoming experiments. In the meantime, if you haven't checked out Gal's blog, you should...

...teams succeeded with this approach around the same time (He et al., 2024; Tong et al., 2024; Ye et al., ...to mitochondrial base editing to base editors optimized for particular species and more, base editors ... X. (2024). Protein language models-assisted optimization of a uracil-N-glycosylase variant enables programmable...

...certain number of days and tracking average transit time for each courier we contracted with so we can always...navigated customs and arrived at their destination in a timely manner and at the correct temperature. To date,...written resources are valuable, there does come a time when a picture is worth a thousand words, especially...

...Addgene repository can save researchers significant time and removes a variable from the lengthy process ... patterns so different? It could be a matter of timing, as jGCaMP7 was released relatively soon after ...informed choice on which AAV to request and/or save time in selecting experimental conditions. Sharing our...

...generated using standard methods that have been optimized for each specific virus in order to generate high...Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing...gene expression and/or function. These data are sometimes reported or posted on the material page for the...

...authors) are currently working on improving and optimizing ARGs, with Marjorie focused on bacterial ARGs...imaging (Fig. 2d). Creating a cell line can be a time-intensive endeavor. That is why we also included...Genomically mined acoustic reporter genes for real-time in vivo monitoring of tumors and tumor-homing bacteria...

... generating mutants. Unexpectedly, phenotypes sometimes differ depending on whether a gene is deactivated... a high-resolution map of early embryogenesis in time and space, and was also used to precisely characterize...redundant) genome, their long three-month generation time and labor-intensive maintenance compared to invertebrate...

...choices. Check out our SpCas9 containing plasmids, optimized for a variety of species.   Specificity concerns...variety of Cas9 containing plasmids that have been optimized for expression in different species and cell types...delivered Cas9 and a sgRNA to your target cells, it is time to confirm that your target sequence has been modified...

...can have low affinity for these tags and even multimeric tags are frequently insufficient for detection...Fluorescent proteins such as GFP have been used for a long time as probes to study connectomics (how neural circuits...microscopy. Sample preparation for these techniques sometimes makes it more difficult to detect protein tags...

...uploading your Sanger sequencing files, either one at a time or as a batch of hundreds, and indicate the guide...do the rest. There are no parameters that need optimizing and no complicated steps to learn. For increased... can compare up to five editing experiments at a time, and batch analysis, which compares hundreds of ...

...to serve as a primer for DNA repair synthesis, ultimately leading to the formation of double Holliday junctions...Challenges: Standardization and HDR CrispyCrunch Optimizing Donor DNA for Enhanced CRISPR Genome Editing ...Methods in Enzymology Vol. 546; (2015). PubMed. Optimization of Genome Engineering Approaches with the CRISPR...

...therapeutic applications where longer retention times often increase therapeutic efficacy (Ahmed et al...re-solubilize and re-fold scFvs from inclusion bodies, it’s time-consuming and laborious.  To avoid this, one or ...https://doi.org/10.3390/antib2020193  Khantasup K, Chantima W, Sangma C, Poomputsa K, Dharakul T (2015) Design...

... videos for free. We’re a small operation, so sometimes it can be a big challenge. But ask any filmmaker...was out the window. So, much like SynBio, it was time to start our Design-Build-Test-Learn cycles. If ...everyone through the recording process was going to be time consuming and difficult. Finding volunteers who ...

...imaging of genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7 (2013): 1479-1491... al. "Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging." Proceedings of...29472486. 17. Adjobo-Hermans, Merel JW, et al. "Real-time visualization of heterotrimeric G protein Gq activation...