Abstract: GPR112 protein, as a member of adhesion class G protein-coupled receptors family (aGPCRs), is encoded by GPR112 gene. It contains seven transmembrane domains and interacts with G protein to perform its functions. Zebrafish is an important model organism for studying the development of vertebrates. Its genome contains the homologous gene GPR1112a of GPR112, which is expressed in both juvenile and adult fish. To study the role of GPR112 in vertebrate survival and development, CRISPR-Cas9 technology was utilized to construct a gpr112a knockout zebrafish line. First, a pair of sgRNAs targeting the second exon of the gpr112a gene were synthesized and microinjected to obtain F0 generation chimeric zebrafish. Next, the F1 generation zebrafish obtained by mating the chimeras with wild-type zebrafish were genotyped. The gpr112a mutant heterozygotes were screened and the gpr112a mutant alleles were subjected to Sanger sequencing to determine the establishment of gpr112a knockout lines. The gpr112a mutant heterozygous zebrafish were then selfed to obtain gpr112a mutant homozygous zebrafish. After microscopic imaging and observation, the zebrafish homozygous for the gpr112a mutation at 7 days post fertilization did not show a phenotype that was significantly different from that of the wild type, which may be caused by the compensatory mechanism of the zebrafish body. This study laid a foundation for exploring the role of GPR112 gene in vertebrate survival and development.
Key words: zebrafish; gpr112a; CRISPR-Cas9; gene knockout; compensatory mechanism
（Acta Laser Biology Sinica, 2022, 31(6): 526-532）