...CRISPR-associated transposases” or MUCICAT. Here, we’ll describe each of these CRISPR systems. The original pCas...by sacB counterselection.  The MUCICAT toolkit includes versions of plasmids with different induction ...mechanisms behind their distinct phenotypes and to design a new generation of engineered cells for green ...

...series of markers are set up sequentially, from broadest to most specific marker, and are represented as...are, try drawing a quadrant over the plot that divides it into four, using the location of the cell populations...Gating and plot orders  As the gates progress from broadest markers to most specific, the first step is to...

... If not, our protocol on Lentivirus Production provides a general approach for producing lentivirus in...excellent protocol paper by the Feng Zhang lab describing construction, lentiviral production, transduction...the Broad Institute- A variety of protocols and guides regarding pooled libraries, from cloning a pooled...

...promising tool for future structure-guided drug design and studies of protein ubiquitination and degradation...CRBNmidi plasmid here!   Kroupova, A. et al. (2024). Design of a Cereblon construct for crystallographic and...specifically for C. elegans (Cao et al., 2024). The team designed coding sequences for mScarlet3 and mScarlet-I3...

...to distinguish from background tissue unless destructive ultrasound imaging was used. We set out to develop...acoustic properties of these gas vesicles made non-destructive specific imaging in tissue difficult. The first-generation... allowed the GVs to be flexible and produce nondestructive nonlinear ultrasound contrast. It turned out...

...other words, your optogenetic experiment should be designed around a behavioral test in which your circuit...of mouse transgenics. Everything that has been described is intended for the mouse; however, all the steps....Diester, Ilka, et al. "An optogenetic toolbox designed for primates." Nature neuroscience 14.3 (2011)...

...human cells (Kim et al, 2016). The Waldman lab has designed their AAV tagging vectors (pAAV-SEPT-Acceptor ...vectors for your specific gene, the lab recommends designing homology arms with the modification of interest...combining the AAV and CRISPR-based approaches is described in several recent reviews (Howes and Schofield...

..., it’s essential to understand them in order to design the most specific assays possible. In this blog...-target effects should impact your experimental design differently. Yes, it is always better to reduce...introducing a specified edit). However, if your design does have wiggle room, you can minimize off targets...

...facilitate the selection of cells containing your desired edit. The selection is marker is then seamlessly...sites is cytotoxic for proliferating cells and provides a simple tool for conditional cell ablation.,”...Zinc-finger nuclease-induced gene repair with oligodeoxynucleotides: wanted and unwanted target locus modifications...

...good starting point for most researchers as it provides a list of putative off-target sites that they ... approach misses larger mismatches (e.g. six nucleotides) that may still lead to off-target double-stranded...contains both the rare off target event and the desired edit is low. Therefore, unbiased sequencing may...

...experiments to minimize it. Overview of DIO/FLEx vector design Recombinase-dependent vectors utilize pairs of ...between them. Early recombinase-dependent vector designs used a lox-STOP-lox (LSL) or frt-STOP-frt (FSF)...you intend to use performs in your system. This includes a thorough testing of both the serotype and promoter...

...tetroxide). These conditions destroy most PA-FPs, but, in 2015, the Looger Lab designed 2 variants of EosFP that...to green). For example, mEosFP (named after the goddess of dawn in Greek mythology) switches from green...

...in the development of CBEs and ABEs. This post includes a lot of acronyms, so we’ve included a glossary... in different cell types, available at www.crisprbehive.design. CGBEs fused to pioneer factors to improve...strand that does not base pair with the guide RNA. Despite being called the "non-target strand" in CRISPR ...

...second edition includes new and updated content in a clean and easy-to-navigate design. You can find it...

...quest, which led us to Seqera Labs, a company that provides open source workflow orchestration software for...on how Addgene has used Nextflow to improve and redesign our pipeline. In it, they break down the unique...

...#12DaysOfCRIPSR hashtag to share new CRISPR content designed to help you with your CRISPR experiments and to...CRISPR, Addgene gave to me: tips and tricks for designing gRNAs for CRISPR editing. On the fourth day of...

...distribution. Lab tip #3: Barcode everything Use barcodes and scanners to keep track of reagents in your...materials quickly and easily. Learn more about using barcodes. Check out our “Lab Tip” filter on the blog...

...anniversary! We've done so much in the past two (2!) decades that it's hard to summarize, but we wanted to mark...experience we're sharing through blogs, eBooks, videos, guides, protocols, and of course, the friendly folks at...

...and find our guides, plasmid kits and other resources for microbial research. Kits, guides, plasmids, ...

...article describing a sucrose supplementation strategy to improve yield. In addition, as described in this...