Fig. 3.

Schematic diagram of the construction of RGS14-KO mice using the CRISPR-Cas9 method and identification of RGS14 expression. a One sgRNA targeting a region downstream of the 3′ end of exon 3 in the RGS14 mouse gene was designed and constructed. b After microinjection, a T7E1 assay indicated that four out of six pups contained cleavage products, suggesting a mixture of mutant and wild-type DNA templates in these mice. c Following subcloning of the PCR products, eight subclones of each mouse were sequenced. All of the subclones carried a single mutant allele, whereas two indels (#5–5, #5–6) produced frameshift mutations. Founder #5–5 was mated to a C57BL/6J mouse to obtain the F1 generation. d Gene sequencing for RGS14 expression levels in hearts from wild-type and RGS14 knockout groups. e Representative western blots for RGS14 expression levels in hearts from the RGS14 ^+/+ and RGS14 ^−/− groups