Utilizing CRISPR technologies to probe Staphylococcus epidermidis' role in the human skin microbiome

Destiny Porter

Document Type

Article

Publication Date

Summer 2022

Keywords

JGM

JAX Location

In: Student Reports, Summer 2022, The Jackson Laboratory

Abstract

The human skin harbors distinct communities of microbes that influence the overall health of the host; one of the most prevalent microbes on the human skin being Staphylococcus epidermidis. S. epidermidis is both an essential commensal skin microbe is one of the most common sources of nosocomial infection. Its ability to survive in various environments on the body, its ability to form dense biofilms, its resistance to commonly used antibiotics makes it of particular interest to researchers. The Oh lab takes particular interest in analyzing S. epidermidis strain diversity and the genes that contribute to this diversity to better understand its role in the human skin microbiome. Utilizing CRISPRi gene knockdown technology, the Oh lab expected to characterize gene function and necessity within different strains of S. epidermidis. However, the use of CRISPRi technology came with limitations that made researching strain diversity in an efficient and cost-effective manner a challenge. As a result of these limitations, a modified CRISPR technology originally created for use in Staphylococcus aureus (CRISPR adaptation-mediated library manufacturing) is being utilized in S. epidermidis to efficiently create libraries of CRISPR RNAs. For this project, I tested the CRISPR adaptation-mediated library manufacturing (CALM) technology in various strains of S. epidermidis to characterize strain dependent differences in CALM efficiency. Additionally, I grew various strains of S. epidermidis to characterize differences in growth attributed to strain diversity and whether CALM machinery is present.

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