![]() However, production and purification of AAV not only represents a significant clinical manufacturing challenge ( Loo and van derWright, 2016 Halbert et al., 2018 Davidsson et al., 2020), it also limits more widespread use of this approach in the research community.īuilding on previous work, including a protocol for CRISPR/Cas9-mediated gene knock-out in human and murine T cells ( Seki and Rutz, 2018 Oh et al., 2019) and a report describing the use of linear dsDNA as repair template ( Roth et al., 2018), we set out to develop a robust, efficient, and scalable protocol for nonviral CRISPR/Cas9-mediated gene knock-in in primary human T cells. Several studies have subsequently reported high editing efficiencies using AAV-based repair templates ( Choi et al., 2019 Vakulskas et al., 2018 Dai et al., 2019). This approach was used, for instance, to insert a CAR construct into the T cell receptor α constant ( TRAC) region locus, which placed the CAR under the control of the endogenous TCR promotor, thus improving its performance ( Eyquem et al., 2017). Viral vectors, in particular adeno-associated viruses (AAV), have been used to deliver donor DNA templates for HDR-mediated target gene knock-in in T cells ( Sather et al., 2015 Wang et al., 2016 Eyquem et al., 2017 Choi et al., 2019). In lieu of that, however, the Lumetri Scopes stands solidly as a surrogate assistant.Homology-directed repair (HDR) of double-strand DNA breaks introduced by targeted gene editing methods, such as transcription activator–like effector nucleases, zinc finger nucleases, or CRISPR/Cas9, can be used to make precise changes to a genomic sequence, including the insertion of long stretches of DNA at a defined genomic location ( Li et al., 2020 Singh et al., 2017). What’s most important is listening to them when they’re telling you that something is awry.Ī second pair of eyes in the editing bay never hurts. As you advance as an editor and as a colorist, you’ll find new ways of implementing each of them into your routine. Which is the best Lumetri Scope? We’ve all got our favorite. ![]() Using the Lumetri Scopes: Color Correction Will Never Be the Same Related: Tips for Editing Videos Like a Pro You’re able to compare the look of all of your footage and adjust each shot to match easily when everything is laid out. The RGB Parade, on the other hand, is great when coloring chromatic tones for continuity. It gives you a very clear reading of your shot’s luminance in a global sense. If it isn’t, this handy guide gives you something tangible to be working these values toward.Īs for the Waveform, this scope will be the first to tell you when a shot is over-exposed or under-exposed. If your footage is well-calibrated, any human skin on the screen should be creeping up along this line, no matter what the actor’s ethnicity happens to be. Picture the big hand of an analog clock at approximately 10:30. The Vectorscope will be especially useful any time you’ve got human beings in-frame. ![]() All of them are useful in their own unique ways.Īnother scenario: skin is one of the most important things to get right as a colorist, and it can be really difficult to nail it by eye. No matter what your preferred configuration of displays is, you’ll likely be using these scopes both for evaluating footage immediately after ingestion or while coloring your project after you’ve locked the picture.Įach scope focuses on a different aspect of coloring footage. This way, you’ll be able to assess each shot technically before exporting. As you prepare to make your final delivery, you’re going to want to make sure that this option is unchecked. You can also toggle the Clamp Signal option on or off to constrain any values lower than 0 and higher than 100 to fall within the gamut of acceptable values. You have a couple of display options aside from these presets, including the ability to choose the color space that you would like to be working with and a choice in terms of the brightness level of the scopes themselves. When two or more scopes are on-screen together, you can rearrange them by double-clicking on any of them. Here, you can see the All Scopes (RGB) preset.
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