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Biology subjects

Friedland, A. E.

Publications and source records attributed to Friedland, A. E..

2 recordsLinked to original sources

Parallel Emergence and Adaptive Evolution of Multicountry Circulating Mycobacterium abscessus Clones

Mycobacterium abscessus (MAB) is a recently emerged bacterial pathogen causing an increasing number of human infections. Despite increasing recognition that different circulating clones contribute disproportionately to MAB population expansion, the global population dynamics and the evolutionary pressures underlying the emergence and spread of MAB clones remain incompletely understood. We analyzed 11,314 publicly available genomes of MAB isolates sampled from 30 countries and identified 38 recently emerged clones that have spread across multiple countries, including the seven previously described Dominant Circulating Clones (DCCs). There was a significant association between the time since the emergence of each circulating clone and its population size, with older clones generally having larger populations. Among the 26 circulating clones of M. abscessus subsp. abscessus, 12 carried the macrolide-susceptible erm(41) 28C genotype, highlighting clone-level variation in macrolide susceptibility. Genome-wide analyses identified 55 genes that were previously under purifying selection but showed evidence of positive selection during recent clade expansion. Collectively, our findings reveal that the recent global expansion of MAB comprises a broad continuum of newly emerged clones, accompanied by widespread shifts in gene-level selective pressures indicative of ongoing adaptation to new ecological environments.

microbiology↗

Pairwise library screen systematically interrogates Staphylococcus aureus Cas9 specificity in human cells

We report a high-throughput screening approach to measure Staphylococcus aureus Cas9 (SaCas9) genome editing variation in human cells across a large repertoire of 88,692 single guide RNAs (sgRNAs) paired with matched or mismatched target sites in a synthetic cassette. We incorporated randomized barcodes that enable whitelisting of correctly synthesized molecules for further downstream analysis, in order to circumvent the limitation of oligonucleotide synthesis errors. We find SaCas9 sgRNAs with a 21-nucleotide spacer are most active against off-targets with single and double mismatches, compared to shorter or longer sgRNAs. Using this dataset, we developed an SaCas9 specificity model that performs well in ranking off-target sites. The barcoded pairwise library screen enabled high-fidelity recovery of guide-target relationships, providing a scalable framework for the investigation of CRISPR enzyme properties and general nucleic acid interactions.

bioengineering↗