Characterization of a thermostable Cas12a ortholog

Wu, Jing; Gao, Pan; Shi, Yajing; Zhang, Caixiang; Tong, Xiaohan; Fan, Huidi; Zhou, Xi; Zhang, Ying; Yin, Hao

doi:10.1016/j.cellin.2023.100126

Cited by 6 publications

(1 citation statement)

References 42 publications

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“…As it currently stands, there is a lack of broadly applicable genetic tools, including selectable markers, promoters, plasmid origins, and transformation protocols, as broadly utilized mesophilic tools are not functional in these thermophilic microbes (Y. Wang et al., 2024 ; Ye et al., 2023 ). While there are a growing number of tools being generated for use in thermophilic microbes (Adalsteinsson et al., 2021 ; Le & Sun, 2022 ; Riley et al., 2019 ; Walker et al., 2020 ; Wang et al., 2022 ; Wu et al., 2023 ; Yang et al., 2023 ), developing new tools and implementing CRISPR mediated genome engineering and/or thermostable serine recombinase-assisted genome engineering (SAGE) mediated integration would greatly simplify testing production of various industrial chemicals in these currently underutilized microbes (Fenster & Eckert, 2021 ; Wu et al., 2023 ). Continued decrease in -omics costs coupled with advanced genome modification techniques makes adopting extremophiles and other novel, less studied microbes as industrial chassis a viable and exciting avenue of research that should be more intensely pursued.…”

Section: Discussionmentioning

confidence: 99%

Application of functional genomics for domestication of novel non-model microbes

Bales,

Vergara,

Eckert

2024

Journal of Industrial Microbiology and Biotechnology

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With the expansion of domesticated microbes producing biomaterials and chemicals to support a growing circular bioeconomy, the variety of waste and sustainable substrates that can support microbial growth and production will also continue to expand. The diversity of these microbes also requires a range of compatible genetic tools to engineer improved robustness and economic viability. As we still do not fully understand the function of many genes in even highly studied model microbes, engineering improved microbial performance requires introducing genome-scale genetic modifications followed by screening or selecting mutants that enhance growth under prohibitive conditions encountered during production. These approaches include adaptive laboratory evolution, random or directed mutagenesis, transposon-mediated gene disruption, or CRISPR interference (CRISPRi). Although any of these approaches may be applicable for identifying engineering targets, here we focus on using CRISPRi to reduce the time required to engineer more robust microbes for industrial applications. One-Sentence Summary The development of genome scale CRISPR-based libraries in new microbes enables discovery of genetic factors linked to desired traits for engineering more robust microbial systems.

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