Altered Splicing in Stable Cell Strains Expressing Mini-hF9 Gene with Nonsense Mutation.
10.19746/j.cnki.issn.1009-2137.2021.01.037
- Author:
Gang WANG
1
;
Wen-Wen SUN
2
;
Lv-Kai ZHU
2
;
Yan-Chun MA
2
;
Xia-Lin ZHANG
2
;
Jian-Hua ZHANG
2
;
Juan REN
2
;
Xiu-Yu QIN
2
;
Lin-Hua YANG
3
;
Bao-Feng CHAI
4
Author Information
1. Department of Hematology, The Second Clinical Medical College (The Second Hospital), Shanxi Medical University,Taiyuan 030001, Shanxi Province, China,Institute of Hematology, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China.
2. Department of Hematology, The Second Clinical Medical College (The Second Hospital), Shanxi Medical University,Taiyuan 030001, Shanxi Province, China.
3. Department of Hematology, The Second Clinical Medical College (The Second Hospital), Shanxi Medical University,Taiyuan 030001, Shanxi Province, China,Institute of Hematology, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China,E-mail: yanglh5282@163.com.
4. Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan 030006, Shanxi Province, China,E-mail: bfchai@sxu.edu.cn.
- Publication Type:Journal Article
- MeSH:
Codon, Nonsense;
Factor IX/metabolism*;
HeLa Cells;
Humans;
Mutation;
RNA Splicing;
RNA, Messenger/metabolism*
- From:
Journal of Experimental Hematology
2021;29(1):228-232
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the molecular mechanism in stable cell strains expressing Mini-hF9 gene with nonsense mutation.
METHODS:Mini-hF9 gene and its nonsense mutants were transfected into HeLa cells independently, and stable cell strains were obtained after G418 resistance screening and monoclonal transformation. The altered splicing and protein expression of mRNA in Mini-hF9 gene in stable cell strains were detected by using RT-PCR and Western blot.
RESULTS:The wild type and nonsense mutated human coagulation factor IX stable cell strains were constructed successfully, which were named HeLa-F9-WT, HeLa-F9-M1 and HeLa-F9-M2. Only normal splicing Norm was detected in the wild-type cell strain HeLa-F9-WT; Norm and Alt-S1 splicing were detected in HeLa-F9-M1; while Norm, Alt-S1 and Alt-S2 splicing were detected in HeLa-F9-M2.
CONCLUSION:The nonsense associated altered splicing (NAS) pathway, which generated alternately spliced transcripts, might be triggered in coagulation factor IX gene with nonsense mutation.
