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CRISPR-Mediated Engineering across the Central Dogma in Plant Biology for Basic Research and Crop Improvement

Molecular Plant 2021年第1期14卷 127-150页

作者： Dibyajyoti Pramanik rahul mahadev shelake Mi Jung Kim Jae-Yean Kim Division of Applied Life Science(BK21 Four Program) Plant Molecular Biology and Biotechnology Research CenterGyeongsang National UniversityJinju 52828Korea These authors contributed equally to this article

The central dogma(CD)of molecular biology is the transfer of genetic information from DNA to RNA to *** CD processes governing genetic flow include the cell cycle,DNA replication,chromosome packaging,epigenetic changes,transcription,posttranscriptional alterations,translation,and posttranslational *** CD processes are tightly regulated in plants to maintain genetic integrity throughout the life cycle and to pass genetic materials to next *** of various CD processes involved in gene regulation will accelerate crop improvement to feed the growing world *** technology enables programmable editing of CD processes to alter DNA,RNA,or protein,which would have been impossible in the ***,an overview of recent advancements in CRISPR tool development and CRISPR-based CD modulations that expedite basic and applied plant research is ***,CRISPR applications in major thriving areas of research,such as gene discovery(allele mining and cryptic gene activation),introgression(de novo domestication and haploid induction),and application of desired traits beneficial to farmers or consumers(biotic/abiotic stress-resilient crops,plant cell factories,and delayed senescence),are ***,the global regulatory policies,challenges,and prospects for CRISPR-mediated crop improvement are discussed.

关键词： central dogma CRISPR crop improvement gene regulation genome editing

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Heterologous Expression of Thermolabile Proteins Enhances Thermotolerance in Escherichia coli

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Advances in Microbiology 2016年第9期6卷 602-612页

作者： Yuya Ueda Seiji Yamauchi Shinsuke Fukata Hidetoshi Okuyama Eugene Hayato Morita rahul mahadev shelake Hidenori Hayashi Graduate School of Science and Engineering Ehime University Matsuyama Japan Proteo-Science Center Ehime University Matsuyama Japan Laboratory of Environmental Molecular Biology Faculty of Environmental Earth Science Hokkaido University Sapporo Japan Faculty of Agriculture Ehime University Matsuyama Japan Department of Chemistry Faculty of Science Josai University Saitama Japan

Heat shock proteins (HSPs) play important roles in the mechanism of cellular protection against various environmental stresses. It is well known that accumulation of misfolded proteins in a cell triggers the HSPs expression in prokaryotes as well as eukaryotes. In this study, we heterologously expressed two proteins in E. coli, namely, citrate synthase (CpCSY) and malate dehydrogenase (CpMDH) from a psychrophilic bacterium Colwellia psychrerythraea 34H (optimal growth temperature 8°C). Our analyses using circular dichromism along with temperature-dependant enzyme activities measured in purified or direct cell extracts confirmed that the CpCSY and CpMDH are thermolabile and present in misfolded form even at physiological growth temperature. We observed that the cellular levels of HSPs, both GroEL and DnaK cheperonins were increased. Similarly, higher levels were observed for sigma factor s32 which is specific to heat-shock protein expression. These results suggest that the misfolded-thermolabile proteins expressed in E. coli induced the heat shock response. Furthermore, heat treatment (53°C) to wild type E. coli noticeably delayed their growth recovery but cells expressing CpCSY and CpMDH recovered their growth much faster than that of wild type E. coli. This reveals that the HSPs expressed in response to misfolded-thermolabile proteins protected E. coli against heat-induced damage. This novel approach may be a useful tool for investigating stress-tolerance mechanisms of E. coli.

关键词： Heat Shock Proteins σ32 Psychrophilic Proteins Thermotolerance

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Engineering drought and salinity tolerance traits in crops through CRISPR-mediated genome editing:Targets,tools,challenges,and perspectives

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Plant Communications 2022年第6期3卷 208-234页

作者： rahul mahadev shelake Ulhas Sopanrao Kadam Ritesh Kumar Dibyajyoti Pramanik Anil Kumar Singh Jae-Yean Kim Division of Applied Life Science(BK21 Four Program) Plant Molecular Biology and Biotechnology Research CenterGyeongsang National UniversityJinju 52828Korea Department of Agronomy&Plant Genetics University of MinnesotaSaint PaulMN 55108USA ICAR-National Institute for Plant Biotechnology LBS CentrePusa CampusNew Delhi 110012India Division of Life Science Gyeongsang National University501 Jinju-daeroJinju 52828Korea

Prolonged periods of drought triggered by climate change hamper plant growth and cause substantial agricultural yield losses every *** addition to drought,salinity is one of the major abiotic stresses that severely affect crop health and agricultural *** responses to drought and salinity involve multiple processes that operate in a spatiotemporal manner,such as stress sensing,perception,epigenetic modifications,transcription,post-transcriptional processing,translation,and post-translational ***,drought and salinity stress tolerance are polygenic traits influenced by genomeenvironment *** of the ideal solutions to these challenges is the development of highyielding crop varieties with enhanced stress tolerance,together with improved agricultural ***,genome-editing technologies,especially clustered regularly interspaced short palindromic repeats(CRISPR)tools,have been effectively applied to elucidate how plants deal with drought and saline *** this work,we aim to portray that the combined use of CRISPR-based genome engineering tools and modern genomic-assisted breeding approaches are gaining momentum in identifying genetic determinants of complex traits for crop *** review provides a synopsis of plant responses to drought and salinity stresses at the morphological,physiological,and molecular *** also highlight recent advances in CRISPR-based tools and their use in understanding the multi-level nature of plant adaptations to drought and salinity *** CRISPR tools with modern breeding approaches is ideal for identifying genetic factors that regulate plant stress-response pathways and for the introgression of beneficial traits to develop stress-resilient crops.

关键词： CRISPR/Cas drought tolerance genome editing polygenic traits salt tolerance trait introgression

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