Journal: Nature Communications

Article Title: CoCas9 is a compact nuclease from the human microbiome for efficient and precise genome editing

doi: 10.1038/s41467-024-47800-9

Figure Lengend Snippet: a Phylogenetic tree of selected Cas9 subfamilies mostly used for genome editing applications (blue) or with experimentally determined three-dimensional structure (orange). Protein alignments: gray, aligned protein sequences; black, conserved residues. Length: number of amino acids. b Scheme of the CRISPR locus where CoCas9 is located. c tracrRNA and crRNA predicted secondary structure. d Sequence logo representation of the PAM of CoCas9, determined using an in vitro cleavage assay. e Heatmap showing relative frequency of the 256 PAM nucleotide combinations at the four most informative positions, compared to the non-cleaved PAM library. Source data are provided as a Source Data file.

Article Snippet: In brief: the synthetic DNA encoding the human codon optimized version of the Cas9 genes was obtained from Genscript and cloned into an expression vector for in vitro transcription and translation (IVT) (pT7-N-His-GST Thermo Fisher Scientific).

Techniques: CRISPR, Sequencing, In Vitro, Cleavage Assay

Journal: Microbial Cell Factories

Article Title: Complete microbial synthesis of crocetin and crocins from glycerol in Escherichia coli

doi: 10.1186/s12934-023-02287-9

Figure Lengend Snippet: Engineered crocin biosynthetic pathway in Escherichia coli

Article Snippet: The E. coli codon-optimized versions of the plant-derived genes (csCCD2 from Crocus sativus , GjUGT1 from Gardenia jasminoides , GT1-316 from Populus fremontii x Populus angustifolia , NtUGT from Nicotiana tabacum , FaGT2 from Fragaria ananassa , StUGT from Solanum tuberosum , CaUGT3 from Catharanthus roseus , NsUGT from Nicotiana sylvestris , and SpUGT from Solanum pennellii ) were chemically synthesized (GenScript, Piscataway, NJ, USA) and individually subcloned into a plasmid pKK223-3 (Table ; Additional file : Table for a sequence of the synthetic genes).

Techniques:

Journal: Microbial Cell Factories

Article Title: Complete microbial synthesis of crocetin and crocins from glycerol in Escherichia coli

doi: 10.1186/s12934-023-02287-9

Figure Lengend Snippet: Microbial crocetin/crocin production

Article Snippet: The E. coli codon-optimized versions of the plant-derived genes (csCCD2 from Crocus sativus , GjUGT1 from Gardenia jasminoides , GT1-316 from Populus fremontii x Populus angustifolia , NtUGT from Nicotiana tabacum , FaGT2 from Fragaria ananassa , StUGT from Solanum tuberosum , CaUGT3 from Catharanthus roseus , NsUGT from Nicotiana sylvestris , and SpUGT from Solanum pennellii ) were chemically synthesized (GenScript, Piscataway, NJ, USA) and individually subcloned into a plasmid pKK223-3 (Table ; Additional file : Table for a sequence of the synthetic genes).

Techniques:

Journal: Microbial Cell Factories

Article Title: Complete microbial synthesis of crocetin and crocins from glycerol in Escherichia coli

doi: 10.1186/s12934-023-02287-9

Figure Lengend Snippet: E. coli strains and plasmids used in this study

Article Snippet: The E. coli codon-optimized versions of the plant-derived genes (csCCD2 from Crocus sativus , GjUGT1 from Gardenia jasminoides , GT1-316 from Populus fremontii x Populus angustifolia , NtUGT from Nicotiana tabacum , FaGT2 from Fragaria ananassa , StUGT from Solanum tuberosum , CaUGT3 from Catharanthus roseus , NsUGT from Nicotiana sylvestris , and SpUGT from Solanum pennellii ) were chemically synthesized (GenScript, Piscataway, NJ, USA) and individually subcloned into a plasmid pKK223-3 (Table ; Additional file : Table for a sequence of the synthetic genes).

Techniques: Plasmid Preparation, Sequencing, Cloning, Expressing