期刊
MOLECULAR BIOLOGY AND EVOLUTION
卷 38, 期 5, 页码 2030-2044出版社
OXFORD UNIV PRESS
DOI: 10.1093/molbev/msab004
关键词
multigene families; functional diversification; transcriptional niche; RNAseq; odorant-binding proteins; evolutionary genetics
资金
- US Department of Education (Graduate Assistance in Areas of National Need [GAANN] fellowship)
- National Institute of General Medical Sciences of the National Institutes of Health [GM059469, GM128974]
In this study, genetic dissection of the insect odorant-binding protein family Obp50 cluster using CRISPR technology revealed redundancy, additivity, diversification, and epistasis among paralogs. Different gene editing combinations had significant effects on organismal phenotypes, indicating functional diversification within the Obp50 cluster. The deletion-reinsertion approach applied in this study provides a general paradigm for dissecting paralogs of multigene families.
Large multigene families, such as the insect odorant-binding proteins (OBPs), are thought to arise through functional diversification after repeated gene duplications. Whereas many OBPs function in chemoreception, members of this family are also expressed in tissues outside chemosensory organs. Paralogs of the Obp50 gene cluster are expressed in metabolic and male reproductive tissues, but their functions and interrelationships remain unknown. Here, we report the genetic dissection of four members of the Obp50 cluster, which are in close physical proximity without intervening genes. We used CRISPR technology to excise the entire cluster while introducing a PhiC31 reintegration site to reinsert constructs in which different combinations of the constituent Obp genes were either intact or rendered inactive. We performed whole transcriptome sequencing and assessed sexually dimorphic changes in transcript abundances (transcriptional niches) associated with each gene-edited genotype. Using this approach, we were able to estimate redundancy, additivity, diversification, and epistasis among Obp50 paralogs. We analyzed the effects of gene editing of this cluster on organismal phenotypes and found a significant skewing of sex ratios attributable to Obp50a, and sex-specific effects on starvation stress resistance attributable to Obp50d. Thus, there is functional diversification within the Obp50 cluster with Obp50a contributing to development and Obp50d to stress resistance. The deletion-reinsertion approach we applied to the Obp50 cluster provides a general paradigm for the genetic dissection of paralogs of multigene families.
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