TL;DR
This work describes how to generate degron knock-ins by CRISPR, cross them to TIR1-expressing strains, and perform depletion experiments, and provides a description of the current methods and highlight alternative approaches.
This protocol covers both single-gRNA (sgRNA) and dual-gRNA directed knockout systems and describes the procedure for constructing an Auxin-Inducible Degron (AID) knock-in-mediated system to induce the rapid degradation of a target protein in cells, thereby investigating its function.
Summary p53, a key tumor suppressor in animals, prevents DNA damage by arresting the cell cycle, activating repair, or inducing cell death. Mutations in p53 are found in over half of human cancers, making it vital for cancer research. To expand p53 research, we engineered Arabidopsis thaliana to express p53 under an in...
A new series of template vectors suitable for employing AID2 technology in prototrophic C. albicans strains, such as clinical isolates and the reference strain SC5314 are reported, and it is demonstrated that the AID2 system also works in Candida auris, albeit less effectively under some conditions.
By integrating genetic conservation with scalable in vivo phenotyping, C. elegans provides a complementary platform for hypothesis generation, pathway validation, and early-stage therapeutic discovery, while its limitations in adaptive immunity and organ complexity should be recognized when translating findings to huma...
In the nucleus, newly synthesized precursor mRNAs undergo several processing steps, such as 5' capping, splicing, polyadenylation and 3' end cleavage, resulting in the formation of an export-competent ribonucleoprotein (mRNP) complex.
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