Advances in DNA sequencing and machine learning are illuminating protein sequences and structures on an enormous scale. However, the energetics driving folding are invisible in these structures and remain largely unknown. The hidden thermodynamics of folding can drive disease, shape protein evolution, and guide protein engineering, and new approaches are needed to reveal these thermodynamics for every sequence and structure.   In this webinar, Dr. Kotaro Tsuboyama presented on cDNA display proteolysis, a new method for measuring thermodynamic folding stability for up to 900,000 protein domains in a one-week experiment. Using this immense dataset, they quantified (1) environmental factors influencing amino acid fitness, (2) the global divergence between evolutionary amino acid usage and protein folding stability and (3) thermodynamic couplings (including unexpected interactions) between protein sites.

First published in 2019, the technique of prime editing offers significant advantages over other genome editing methods, such as CRISPR/Cas9. One critical benefit is the introduction of single-strand, rather than double-strand breaks in the target DNA, which allows the more accurate mismatch repair mechanism to be deployed — reducing the number of unwanted and erroneous edits. In addition, prime editing can install a wider range of edits, including transitions (interchanges of purines or of pyrimidines) and transversions (purines to pyrimidines, and vice versa), as well as deletions and insertions.  Despite its promise, the factors that govern the successful integration of short sequences to a genomic target using prime editing remain unclear. 在本次网络研讨会中，来自英国 Wellcome Sanger 研究所的研究人员将描述他们在不同基因组背景下分析成千上万个插入序列，以了解和预测先导编辑插入效率的开创性工作。结果为获得该结果的机构专有，可能无法反映在其他机构可实现的结果。仅供研究使用。

We find ourselves at an inflection point in functional genomics, one characterized by precision, expanding functionality, and high-throughput capabilities. With this comes a need for high performance tools and experts in the field to help researchers navigate the changing technological landscape and create successful experiments. This eBook offers a contemporary synthesis of CRISPR screening technologies for functional Geneticists, as well as advice from our CRISPR experts.