...your target base in the protospacer sequence. If you do not find a suitable ‘NGG’ PAM at your target genomic...

...saturation mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis? How...

... extension can usually ensure that the 3’-base(s) do not form secondary structures. The introduction (...

...clinical grade hiPSCs, reprogramming methods that do not involve modification of the original genome by...

...find a positive one. So if you have better things to do than clone picking, choose your guide RNAs wisely...

...caption courtesy of Challis et al., 2022. Where do they come from? Systemic capsids are typically derived...efficacy can vary depending on species. LY6A and LY6C1 do not have known homologs in NHPs, and capsids that...mouse strains). AAV-PHP.eC, AAV9-X1.1 and AAV1-X1 do not display strain-specific tropism and can be used...2023). High viral titer High vector genome (vg) doses are recommended for all systemic capsid applications... AAV-PHP.eB is typically administered at a total dose between 1 x 1011 and 5 x 1011 vg per animal (i.e...mL titer), while AAV-PHP.S is typically used at a dose between 3 x 1011 and 1 x 1012 vg*. Most systemic...capsids are easy to produce in high titer, but higher doses (>1 x 1012 vg total in mice) may increase the chances...

...processes in different cell types. One great way to do this is to use CRY2-CIB1 clustering. Fuse either ...provided CIB constructs for targeting Golgi, ER, and endosome membranes) (Figure 4). The authors used SuperPLD...

..., phosphomimetic mutations on the activation loop do not fully activate MEK1. In fact, replacing just ...

...comparison. These next-generation sequencing techniques do not typically rely on Sanger sequencing but instead...

...negative selection markers for allelic exchange. To do this, we performed the following modifications to...

...utmost priority for countries around the world and to do so, the WHO recommends one strategy: testing, tracking...Servellita V, Singh J, Miao X, Streithorst JA, Granados A, Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu ...

... by restriction enzyme cloning, you would have to do it in two steps, and a scar would likely remain between...

...default state. In contrast, Brainbow-3.1 and -3.2 do not display default fluorescence due to the addition...

...systems listed above are orthogonal, meaning that they do not affect each other. For example, the GAL4 transcription...

...near optimal wavelengths (Fluc emits at 610 nm) and do not require external light sources. Unfortunately...

...you’ve introduced your RNAi into your organism, how do you identify the actively silenced ones? Here, I’...

...recombinant clones are generated.  A simple way to do this is to express lambda red genes from a plasmid...

...success, we find it is usually faster and easier to do the validation first.  AAV-mediated gene expression...

...μm) tissue sections and 3D cultures These samples do not necessarily require optical sectioning, but open...while also avoiding the toxic effects of high light doses to the cells. Thin static samples Ex: Fixed monolayers...out of focus light. However, they also continually dose the sample from top to bottom with excitation light...

... a new or experienced reprogrammer? Which methods do you have experience using? Let us know about your...multiple transfection makes it difficult to control the dose of plasmid the cells receive over the whole reprogramming...