中国输血杂志2023，Vol.36Issue(11)：986-990,5.DOI:10.13303/j.cjbt.issn.1004-549x.2023.11.003
中国人群中RHAG变异对mRNA剪接影响的体外研究
Effect of RHAG variants identified in Chinese population on RHAG mRNA splicing in vitro
摘要
Abstract
Objective To study the effect of RHAG variants identified in Chinese population on mRNA splicing by mini-gene splicing assay(MSA)in vitro.Methods The pSplice POLR2G minigene expression plasmids were constructed for 10 RHAG mutations with relatively high distribution frequency in Chinese population near splicing sites or synonymous mutations by analyzing the RHAG gene data in the KMxD database.Then,the wild-type and mutant plasmids were transfected into HEK 293T cells,and RNA was extracted 48 hours after transfection.After reverse transcription,specific primers were used for PCR amplification,and then agarose gel electrophoresis and capillary electrophoresis were performed to determine whether the mutations will affect the normal splicing of exons.Results MSA in vitro showed that 2 mutations(c.158-5delT,c.807+3A＞C)near the splicing site reduced the amount of normal transcripts slightly.The remaining 8 synonymous mutations(c.312G＞A,c.341+3G＞A,c.609C＞T,c.681G＞A,c.861G＞A,c.957T＞A,c.984T＞C and c.1139-7G＞A)had no impact on the splicing of RHAG mRNA.Conclusion This study showed that RHAG gene was conservative in terms of splicing,and the mutations near splicing sites and synonymous mutations were less likely to cause abnormal splicing of RHAG gene.关键词
RHAG突变/体外微基因剪接系统/剪接异常/中国人群Key words
RHAG variants/minigene splicing assay(MSA)/aberrant splicing/Chinese population分类
医药卫生引用本文复制引用
贾双双,孙明明,温机智,魏玲,罗广平,姬艳丽..中国人群中RHAG变异对mRNA剪接影响的体外研究[J].中国输血杂志,2023,36(11):986-990,5.基金项目
广东省基础与应用基础研究基金项目(2021A1515012139) （2021A1515012139）
广州市卫生健康科技项目(20231A010047) （20231A010047）
广州市科技计划项目(2023A03J0552) （2023A03J0552）
广州市医学重点学科(2021-2023)项目 （2021-2023）
