Glycosyl residue composition of cell wall extracts from tillers of switchgrass WT and PvGAUT4-KD plants
Glycosyl residue composition of cell wall extracts from tillers of switchgrass WT and PvGAUT4-KD plants
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Engineering topics
Stewart, Neal C.
Publications and source records attributed to Stewart, Neal C..
Glycosyl residue composition and total carbohydrate in cell wall extracts from WT and PvGAUT4-KD lines
Glycosyl residue composition and total carbohydrate in cell wall extracts from WT and PvGAUT4-KD lines
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Glycosyl residue composition of cell wall extracts from stems of P. deltoides WT, vector control and PdGAUT4-KD plants
Glycosyl residue composition of cell wall extracts from stems of P. deltoides WT, vector control and PdGAUT4-KD plants
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Glycosyl residue composition and total carbohydrate of cell wall extracts from WT and PdGAUT4-KD lines
Glycosyl residue composition and total carbohydrate of cell wall extracts from WT and PdGAUT4-KD lines
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Sugar release Supplmentary Table and notes
Sugar release and growth of biofuel crops are improved by downregulation of pectin biosynthesis
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Life Sciences Reporting summary sugar release
Nature Research reporting summary for manuscript
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Sugar Relese Supplementary Text and Figures
Phylogenetic tree of GAUT Protein Family and gene model, RNAi construct, and relative transcript abundance of GAUT4 in switchgrass, rice and poplar knockdown (KD) lines.
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Glycosyl linkage analysis of fractionated cell walls from switchgrass WT and PvGAUT4-KD lines
Glycosyl linkage analysis of fractionated cell walls from switchgrass WT and PvGAUT4-KD lines
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Glycosyl linkage analysis of fractionated cell walls from P.deltoides WT and PdGAUT4-KD lines.
Glycosyl linkage analysis of fractionated cell walls from P.deltoides WT and PdGAUT4-KD lines.
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Epigenetic Footprints of CRISPR/Cas9-Mediated Genome Editing in Plants
CRISPR/Cas9 has been widely applied to various plant species accelerating the pace of plant genome editing and precision breeding in crops. Unintended effects beyond off-target nucleotide mutations are still somewhat unexplored. We investigated the degree and patterns of epigenetic changes after gene editing. We examined changes in DNA methylation in genome-edited promoters of naturally hypermethylated genes (AT1G72350 and AT1G09970) and hypomethylated genes (AT3G17320 and AT5G28770) from Arabidopsis. Transgenic plants were developed via Agrobacterium-mediated floral dip transformation. Homozygous edited lines were selected from segregated T2 plants by an in vitro digestion assay using ribonucleoprotein complex. Bisulfite sequencing comparisons were made between paired groups of edited and non-edited plants to identify changes in DNA methylation of the targeted loci. We found that directed mutagenesis via CRISPR/Cas9 resulted in no unintended morphological or epigenetic alterations. Phenotypes of wild-type, transgenic empty vector, and transgenic edited plants were similar. Epigenetic profiles revealed that methylation patterns of promoter regions flanking target sequences were identical among wild-type, transgenic empty vector, and transgenic edited plants. There was no effect of mutation type on epigenetic status. We also evaluated off-target mutagenesis effects in the edited plants. Potential off-target sites containing up to 4-bp mismatch of each target were sequenced. No off-target mutations were detected in candidate sites. Our results showed that CRISPR/Cas9 did not leave an epigenetic footprint on either the immediate gene-edited DNA and flanking DNA or introduce off-target mutations.
59 BASIC BIOLOGICAL SCIENCES↗