High-Efficiency Genome Editing of Streptomyces Species by an Engineered CRISPR/Cas System.

Next-generation sequencing technologies have rapidly expanded the genomic information of numerous organisms and revealed a rich reservoir of natural product gene clusters from microbial genomes, especially from Streptomyces, the largest genus of known actinobacteria at present. However, genetic engineering of these bacteria is often time consuming and labor intensive, if even possible. In this chapter, we describe the design and construction of pCRISPomyces, an engineered Type II CRISPR/Cas system, for targeted multiplex gene deletions in Streptomyces lividans, Streptomyces albus, and Streptomyces viridochromogenes with editing efficiency ranging from 70% to 100%. We demonstrate pCRISPomyces as a powerful tool for genome editing in Streptomyces.

Effects of color and pattern on implicit and explicit picture memory.

The degree to which repetition priming is perceptually specific is informative about the mechanisms of implicit memory as well as of perceptual processing. In 2 sets of experiments with pictures as stimuli, we tested the effects of color and pattern manipulations between study and test on implicit memory (i.e., naming facilitation) and explicit memory (i.e., 2 forms of recognition). These manipulations did not affect priming. However, participants were able to explicitly detect stimulus changes at above-chance levels. Changes in color also produced small decrements in participants' ability to judge that repeated stimuli were old on a recognition test. Experiment 2 showed diminished priming with changes in the stimulus exemplar (i.e., a different picture of the same named object) from study to test, which demonstrated that the picture-naming paradigm is sensitive to changes in physical attributes. The results suggest that physical attributes that are not essential to the formation of a shape representation do not influence repetition priming in a basic identification paradigm. Suggestions for how priming may be mediated are discussed.