...amplification) or reverse transcriptase (RT)-RPA, respectively. RPA is coupled with T7 transcription to convert amplified...2018) CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science 360...

...used in research, and data repositories. Below I describe how each of these kinds of connectivity facilitates...deposits will be made. The Dataverse integration is described in this video and the Figshare integration in ...from RSpace documents to external data sources (described above) further enhances the value the repository...

...restriction endonucleases in plasmid addition was first described by studies of EcoRI and Bsp6I in Escherichia coli...systems. As with the other toxin-antitoxin systems described above, one part of the pair is more stable than...toxin-antitoxin system discovered. Unlike the systems just described, the hok/sok systems consist of three components...

...heterologously by bacteria inside the body. So we screened gas vesicles gene clusters from a diverse set ...expression heterogeneity required single cell screening to isolate a clonal cell line capable of producing...improvement in expression. Stabilizing the gvpA transcript with WPRE reduced the number of copies of gvpA...

...Cas13a as a diagnostic tool, as discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a   ...Recombinase Polymerase Amplification (RPA) or Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides... the collateral RNA cleavage response is well described in bacterial cells, LwaCas13a does not display...

... The one-pot saturation mutagenesis technique described in Wrenbeck et al is a PCR-based approach for ...library that’s ready to be tested in a functional screen of your liking. This method is similar to site-directed... screen...

...methyltransferase — that can drive lasting transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration...versions using dCas9, zinc finger proteins, and transcription activator-like effectors (TALEs), with fifteen...Weissman, J. S. (2021). Genome-wide programmable transcriptional memory by CRISPR-based epigenome editing. Cell...

...strong transcriptional terminator BBa_B1002 (from IGEM) upstream of the MCS to reduce transcriptional readthrough...worked too. Rhamnose induction relies on the transcriptional activator rhaS. To allow manipulation of bacteria...

...same fluorescent markers, Brainbow 3.1 and 3.2 (described in the same publication) are unique because by...cell type. The Confetti mouse was first used to describe the self-renewal of cells in the intestinal crypt...previous Brainbow technologies were useful for discriminating neurons within only one field of view. Tetbow...

...Easi-CRISPR in vitro transcribes RNA encoding your repair template, then uses reverse transcriptase to generate..., and ultraviolet light. In this post, we will describe the mechanism of homology directed repair, one...

...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool...context-specific effects of UAG suppression, as described (6). To determine the stoichiometry of phospho-incorporated...phospho:non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid...

...applications, including transgenesis, gene-trap screens, and gene editing. How does the piggyBac™ system...effects, such as with signaling molecules or transcription factors, or when the gene is constitutively-..., since there is no selection, you will need to screen somewhere between one hundred and one thousand ...

...Antibody-independent methods may include analysis of transcriptomic or proteomic data from several different tissues... feasible and (2) large-scale proteomic and transcriptomic datasets already exist.   Example: You want...tissues. You know that there is a wealth of transcriptomic data from this sample type, so you use your...

...brightness of individual fluorophores. Table 1 describes the brightness of a few commonly used fluorophores...brightness levels on your smartphone or computer screen), and you should be able to properly label many...panel! Designing a Panel In this example, I will describe a basic panel (Table 2, Figure 5) for a hypothetical...

... drug discovery by big pharma (i.e. HTRF and LanthaScreen) (Degorce et al., 2009), measuring intracellular...are critical in setting up a FRET-based assay or screen (the Addgene blog has an excellent primer on FRET...it all together The FRET donors and acceptors described above can be paired together to measure a wide...

... by abundant transcripts, leading to falsely lowered levels for less abundant transcripts. However, adding...can even be used to detect a single copy of a transcript. But this sensitivity can sometimes cause as ...

...various CRISPR applications (such as visualize, screen, activate, and repress) on Thursday. We’d also ...from nickases to base editing to pooled library screening. It also has information for planning your CRISPR...

...Noninvasive assessment of gut function using transcriptional recording sentinel cells! 🎵 On the 11th day...gave to me: CRISPR activation and interference screens for primary human T cells🎵 On the 3rd day of CRISPR...

...populate your “PubMed filter list” to the left of your screen. That’s it! Next time you log-in to your PubMed...Scientific Review Article Tips for Improving Your Next Manuscript Resources on Addgene.org Learn How to Order...

...used to increase or decrease expression of a gene. Screen: A library of gRNAs with a CRISPR nuclease target...cell population. The resulting mutant cells are screened for phenotypes of interest. Limitations of CRISPR...