TY - JOUR ID - 6003 TI - Experimental validation of a predicted microRNA within human FVIII gene JO - Molecular Biology Research Communications JA - MBRC LA - en SN - 2322-181X AU - Jalali-Qomi, Shahabeddin AU - Motovali-Bashi, Majid AU - Rezaei, Halimeh AU - Khalilian, Sheyda AD - Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Iran Y1 - 2021 PY - 2021 VL - 10 IS - 2 SP - 45 EP - 53 KW - Hemophilia A KW - Factor VIII KW - Small non-coding RNA KW - miRNAs KW - HEK 293 cell line DO - 10.22099/mbrc.2021.39067.1573 N2 - Hemophilia A is an X-linked bleeding disorder that occurs due to the deficiency of Factor VIII (FVIII) protein clotting activity. The mutations in the F8 gene, which encodes FVIII coagulating protein have been widely reviewed. However, there is a wide range of criteria that in addition to F8 gene mutations, different molecular mechanisms may be associated with hemophilia A. Various functions of FVIII could be related to the hypothetical small non-coding RNAs, located within the F8 gene sequence. Therefore, miRNAs that can post-transcriptionally regulate gene expression might confer susceptibility to developing hemophilia A. Here, we have selected a bioinformatically predicted hairpin structure sequence in the first intron of the F8 gene that has the potential to produce a real miRNA (named put-miR1). We tried to experimentally detect the predicted miRNA via RT-PCR following its precursor overexpression in HEK 293 cell lines. Despite the accuracy of miRNA prediction, according to the reliable bioinformatics studies, we couldn’t confirm the existence of considered mature miRNA in transfected cells. We hope that through changing experimental conditions, designing new primers, or altering cell lines and expression vectors, the exogenous and endogenous expression of the predicted miRNA will be confirmed. UR - https://mbrc.shirazu.ac.ir/article_6003.html L1 - https://mbrc.shirazu.ac.ir/article_6003_b3a6da6c068767d97cb6a6bd57981fbf.pdf ER -