Mettl9-/- mice PBMCs
Mus musculus
strain
C57BL/6J
isolate
PBMCs were isolated from mice using Pancoll density gradient medium (Solarbio), washed three times with PBS, harvested, and lysed in gDNA lysis buffer (200 mM NaCl, 100 mM Tris-HCl pH 8.5, 50 mM EDTA pH 8.0, 0.5% SDS, 0.1 mg/mL proteinase K) at 55 C until the solution became clear. Lysates were treated with 0.5 mg/mL RNase A at 37 C for 1 hour to remove RNA. gDNA was then extracted using phenol:chloroform:isopentanol in a 25:24:1 ratio, precipitated with isopropanol, and resuspended in 70% ethanol. The DNA pellet was centrifuged and dissolved in TE buffer.
age
6 to 8 weeks
collection_date
2022-10
geo_loc_name
China
sex
female
tissue
peripheral blood mononuclear cells
treatment
exon 2 of the Mettl9 gene was targeted for knockout using the CRISPR-Cas9 system. Synthesized sgRNA and Cas9 protein were co-injected into fertilized C57BL/6J embryos, which were subsequently implanted to produce F0 mice with the desired gene alteration. These F0 mice were then crossed with C57BL/6J mice to establish a stable F1 line, yielding Mettl9-KO (Mettl9/) mice
BioSampleModel
Model organism or animal