Case Study 1: CRISPR-Cas9 Technology to Knockout the ADE1 Gene in Yeast
Synbio Technologies used CRISPR-Cas9 technology to edit the 920bp ADE1 gene in yeast. ADE1 is a gene involved in adenine synthesis, and its inactivation causes the accumulation of red pigments, resulting in red-colored colonies.
Positive Clone Colonies
Sequencing results showed that the ADE1 gene in the transformants was successfully edited with an 18-base pair deletion.
Case Study 2: CRISPR-Cas9 Technology to Knock-in a Fluorescent Protein Gene in E.coli
Synbio Technologies researchers developed a refined CRISPR-Cas9 dual-plasmid gene-editing system and its production process, successfully knocking in a 906bp mScarlet red fluorescent protein gene into the E. coli NC101 genome at the Clbp coding region. The knock-in success rate ranged from 96.6%~100%.
Figure 1: PCR Identification Electrophoresis of Clones after mScarlet Gene Knock-in
Figure 2: Genomic Sequence of E. coli after mScarlet Knock-in
(a) Gene map and sequencing assembly diagram of upstream and downstream of knock-in site;
(b) Peak shape diagram of upstream and downstream sequencing sequence of knock-in site.
Figure 3: Fluorescence Microscopy Images of E. coli NC101 Mutant Strain
(a) Cell image under white light;
(b) Cell image after excitation with 594 nm light.
DNA Synthesis
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