Figure 7.

CriPs-mediated Nrip1 deletion in white adipocytes increases the expression of UCP1. Primary pre-adipocytes were treated with CriPs with each of four different sgRNAs targeting Nrip1 (Nrip1 sgRNA 1, Nrip1 sgRNA 2, Nrip1 sgRNA 3, and Nrip1 sgRNA 4) and controls. Pre-adipocytes were then differentiated into mature white adipocytes (Cas9–sgRNA, 100 nm; EP, 25 μm). A, indel detection in Nrip1 genomic DNA was determined by a T7E1 assay. DNA bands were as follows: Nrip1 sgRNA 1: uncut, 429 bp; cut, 283 bp + 146 bp; Nrip1 sgRNA 2: uncut, 334 bp; cut, 223 bp + 111 bp; Nrip1 sgRNA 3: uncut, 420 bp; cut, 150 bp + 270 bp; Nrip1 sgRNA 4: uncut, 381 bp; cut, 307 bp + 74 bp. B, UCP1 expression was measured by RT-PCR. C–E, with the treatment of Cas9–Nrip1 sgRNA 3–EP and Cas9–control sgRNA–EP, gene expression was measured by RT-PCR. C, thermogenic genes; D, inflammatory genes; E, neurotrophic factors. Error bars, S.E.