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Nucleic Acids Res 2021 Apr 13. Epub 2021 Apr 13.

Institute of Molecular Infection Biology (IMIB), University of Würzburg, D-97080 Würzburg, Germany.

Antisense peptide nucleic acids (PNAs) inhibiting mRNAs of essential genes provide a straight-forward way to repurpose our knowledge of bacterial regulatory RNAs for development of programmable species-specific antibiotics. While there is ample proof of PNA efficacy, their target selectivity and impact on bacterial physiology are poorly understood. Moreover, while antibacterial PNAs are typically designed to block mRNA translation, effects on target mRNA levels are not well-investigated. Read More

Methods Mol Biol 2021 ;2307:123-137

Chemical and Environmental Engineering, University of California Riverside, Riverside, CA, USA.

Genome-wide functional genomic screens are essential to determining the genetic underpinning of a biological process. Novel and powerful tools for perturbing gene function, with the help of genetic and epigenetic information, have made it possible to systematically investigate the contribution of every gene to evolved and engineered phenotypes. Functional genomics and screening for enhanced phenotypes become ever more important when dealing with nonconventional hosts. Read More

Methods Mol Biol 2021 ;2307:95-109

Chemical and Environmental Engineering, University of California Riverside, Riverside, CA, USA.

Recent developments in RNA-guided nuclease technologies have advanced the engineering of a wide range of organisms, including the nonconventional yeast Yarrowia lipolytica. Y. lipolytica has been the focus of a range of synthetic biology and metabolic engineering studies due to its high capacity to synthesize and accumulate intracellular lipids. Read More

Methods Mol Biol 2021 ;2307:69-83

Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC, USA.

Metabolic engineering frequently requires both gene knockouts and gene integration. CRISPR-Cas9 has been extensively used to create double-stranded DNA breaks that result in indel mutations; however, such mutations can revert or create toxic product. Gene integration can also be accomplished by CRISPR-Cas9 introduced double-stranded DNA breaks and a donor DNA cassette. Read More

Nat Commun 2021 04 12;12(1):2165. Epub 2021 Apr 12.

Department of Genetics, Stanford University, Stanford, CA, USA.

Adenosine-to-inosine (A-to-I) RNA editing catalyzed by ADAR enzymes occurs in double-stranded RNAs. Despite a compelling need towards predictive understanding of natural and engineered editing events, how the RNA sequence and structure determine the editing efficiency and specificity (i.e. Read More