Journal: Cell Genomics

Article Title: Strategic targeting of Cas9 nickase induces large segmental duplications

doi: 10.1016/j.xgen.2024.100610

Figure Lengend Snippet:

Article Snippet: To construct the expression plasmid for nCas9 and gRNAs, we inserted synthetic DNAs encoding gRNAs targeting up10 and down12 into the BbsI and BsaI sites of Addgene plasmid #74630, respectively.

Techniques: Produced, Purification, Virus, Recombinant, DNA Extraction, Plasmid Preparation, Western Blot, Labeling, Membrane, Hybridization, Ligation, Sequencing, Software

Journal: FEBS letters

Article Title: Domain compatibility in Ire1 kinase is critical for the Unfolded Protein Response

doi: 10.1016/j.febslet.2010.06.003

Figure Lengend Snippet: (A) UPR reporter gene assay. AWY14 or AWY19 strains harboring the indicated plasmids were cultured in medium containing galactose for 4 hr followed by addition of β-ME for 4 hr. The activities were reported as Mean ± SEM (n=3). ** and *** represent significant difference (P< 0.05) compared to recombinant Ire1 activity under the same conditions by one-way ANOVA. (B) Relative expression of chimeric IRE1 compared to ACT1 mRNA and efficiency of HAC1 mRNA splicing was analyzed by Scion Image program. HAC1 mRNA splicing upon 2µg/ml Tunicamycin (Tm) was shown in (C). C: RT-PCR without reverse transcriptase. HAC1u and HAC1s represent unspliced and spliced HAC1. M: 1 kb plus DNA ladder.

Article Snippet: Partial fragments of the IRE1 homologs were PCR amplified from genomic DNA of various yeast species using Vent DNA polymerase (New England Biolabs) and Degenerate F1 and Degenerate R1 primers designed from the amino acid sequence of the S. cerevisiae Ire1 kinase in subdomain I and VI ( ).

Techniques: Reporter Gene Assay, Cell Culture, Recombinant, Activity Assay, Expressing, Reverse Transcription Polymerase Chain Reaction