Journal: Cell reports

Article Title: SMYD5 is a regulator of the mild hypothermia response.

doi: 10.1016/j.celrep.2024.114554

Figure Lengend Snippet: Figure 2. Genome-scale CRISPR-Cas9 KO screen on SP1-MHI reveals multiple potential inhibitors and activators of the MHR (A) Overview of the genome-scale CRISPR-Cas9 KO approach for the HEK293WT+Cas9+SP1 cell line. (B) Fluorescence measurements and sort gates of the 4 replicates of transduced HEK293WT+Cas9+SP1 cells (green), negative control (HEK293WT, black) and positive control (HEK293WT+Cas9+SP1, gray). (C and D) Genes marked in red are transcription regulators that have a log10(RRA) score >3.5 and a known repressive function. Genes marked in green are transcription regulators that have a known activating function and a log10(RRA) score >3.5. Colored dots represent genes that have a log10(RRA) score >3.5. Orange dots indicate genes that have a positive log fold change (LFC), and blue dots indicate genes that have a negative LFC, from either the SP1 activator (C) or repressor (D) screen. Log10(RRA) score and LFC were calculated with MAGeCK.29

Article Snippet: These cell lines were made by transfecting the plasmids with lipofectamine as described above and selecting stably transfected cells using Blasticidin (Thermo Fischer Scientific, A1113903) selection for Cas9 and Neomycin(G418/Geneticin) (Santa-Cruz, sc-29065A) selection for SP1-MHI.

Techniques: CRISPR, Fluorescence, Negative Control, Positive Control