... absence of genomic alterations. These studies utilize a catalytically inactive form of Cas9 known as ...elements (CAPTURE) (Liu et al., 2017). This system utilizes dCas9 fused to a biotinylation tag expressed on...

... the reduced requirements for biocontainment precautions, these systems allow researchers to investigate...reconstitute a functional viral polymerase complex and utilize the minigenome as a template. Thus, it is important...

...has occurred. Although the original Y2H systems utilized the yeast Gal4 activator, bacterial LexA DBD and...bait and prey protein fusions to demonstrate the utility of the Y2H system, it is more common to use many...

...track all of our 40,000 samples available for distribution!    Check Out How We Think about Developing...The preferred E. coli strains for storage and distribution at Addgene are DH5α, NEB stable, ccdb survival...

...decided this protocol would be a worthy first contribution to the protocols.io community. Did I mention...scientists workflow in mind and can offer a one-stop solution to your lab's organization and management needs...

...recombine DNA? There are many methods that have been utilized over the years to move around pieces of DNA. Oftentimes...in the DNA. In restriction cloning, scientists utilize specific restriction enzymes to cut dsDNA of interest...reducing the amount of time required for cloning. To utilize this approach, the fragment of DNA that you would...pairs adenine (A) and thymine (T). TOPO® cloning utilizes the Taq polymerase which naturally leaves a single...traditional restriction enzyme cloning. Scientists have utilized this approach to create compatible custom overhangs... 100kb) compared to other cloning methods that utilize bacteria where it becomes progressively more difficult...

...opened up vast possibilities for the clinical and therapeutic applications of iPSCs. The generation of human...the production of iPSCs: Blood is easily and routinely drawn from patients in contrast to the process...

...Cas9. Unfortunately, certain drawbacks limit the utility of these systems: St1Cas9 requires a very specific...  Wang D, Zhang F, Gao G (2020) CRISPR-Based Therapeutic Genome Editing: Strategies and In Vivo Delivery...

...consists of a mutated mCherry and a downstream constitutively active eGFP. To create the inactive mCherry...T2A site and expressed in cells. mCherry is constitutively expressed to identify successfully transfected...

...pathogen or virus detection), therapeutics and sensitive genotyping. The beauty of SHERLOCK systems is that...bottleneck. In the last few years, scientists have utilized CRISPR-Cas9 protein variants, Cas13, and Cas12a...

...invaluable to me as a bench scientist working at an institution where training opportunities weren’t always available...transfer, which occurs when bacteria of different evolutionary lineages share genes that can improve chances...

... processing in live cells. Indeed, groups have utilized dCas13 fusion proteins for imaging, tracking, ...global RNA expression, cell stress, and RNA size distribution. When comparing LwaCas13a to shRNA, they found...

...molecules. Using a technique called systematic evolution of ligands by exponential enrichment (SELEX), ...Peixuan AND Haque, Farzin. RNA Nanotechnology and Therapeutics. Boca Raton: CRC Press, 2014. 5. Hayat, Akhtar...

... spatiotemporal control over its expression. Constitutive or inducible Cre recombinases can be introduced...method to dissect the individual and shared contributions of transcript isoforms to neuronal physiology...

...penetration depth as well as high spatial and temporal resolution, making it ideal for visualizing gene expression...also included an analogous set of transient constitutively-expressed mARG plasmids driven by CMV promoters...

...generation of base modifiers available. This system utilizes an RNA template containing the desired edit adjoined...like with base editing, alternative Cas9s can be utilized allowing for PAM flexibility. Fig. 2: Prime...

... developed, though, researchers at HuidaGene Therapeutics guessed that fusing MPG to an ABE would allow...recognize T and/or C substrates. The HuidaGene Therapeutics team has deposited their editors, called gTBE...

...known functions has the potential to be used in therapeutics, as it can increase gene expression at certain..., Garnett MJ (2019) Prioritization of cancer therapeutic targets using CRISPR–Cas9 screens. Nature 568...

.... An internal control may be a plasmid that constitutively expresses a reporter protein (e.g., GFP or ...many variables. However daunting designing and executing these steps may be, proper controls are a basic...

... Used since the mid-90s, this technique has revolutionised the way we apprehend molecular complexes and...wavelength, to the concentration of fluorophore in solution) are two additional parameters that can affect...