Designing Tyrosinase siRNAs by Multiple Prediction Algorithms and Evaluation of Their Anti-Melanogenic Effects.
10.4062/biomolther.2017.115
- Author:
Ok Seon KWON
1
;
Soo Jung KWON
;
Jin Sang KIM
;
Gunbong LEE
;
Han Joo MAENG
;
Jeongmi LEE
;
Gwi Seo HWANG
;
Hyuk Jin CHA
;
Kwang Hoon CHUN
Author Information
1. Department of Life Sciences, Sogang University, Seoul 04107, Republic of Korea. hjcha@sogang.ac.kr
- Publication Type:Original Article
- Keywords:
Tyrosinase;
Melanin;
siRNA;
Melanocytes;
Whitening
- MeSH:
Dihydroxyphenylalanine;
Gene Expression;
Humans;
Melanins;
Melanocytes;
Melanoma;
Monophenol Monooxygenase*;
RNA;
RNA Interference;
RNA, Small Interfering*;
Skin Pigmentation;
Tyrosine;
Weights and Measures
- From:Biomolecules & Therapeutics
2018;26(3):282-289
- CountryRepublic of Korea
- Language:English
-
Abstract:
Melanin is a pigment produced from tyrosine in melanocytes. Although melanin has a protective role against UVB radiation-induced damage, it is also associated with the development of melanoma and darker skin tone. Tyrosinase is a key enzyme in melanin synthesis, which regulates the rate-limiting step during conversion of tyrosine into DOPA and dopaquinone. To develop effective RNA interference therapeutics, we designed a melanin siRNA pool by applying multiple prediction programs to reduce human tyrosinase levels. First, 272 siRNAs passed the target accessibility evaluation using the RNAxs program. Then we selected 34 siRNA sequences with ΔG ≥−34.6 kcal/mol, i-Score value ≥65, and siRNA scales score ≤30. siRNAs were designed as 19-bp RNA duplexes with an asymmetric 3′ overhang at the 3′ end of the antisense strand. We tested if these siRNAs effectively reduced tyrosinase gene expression using qRT-PCR and found that 17 siRNA sequences were more effective than commercially available siRNA. Three siRNAs further tested showed an effective visual color change in MNT-1 human cells without cytotoxic effects, indicating these sequences are anti-melanogenic. Our study revealed that human tyrosinase siRNAs could be efficiently designed using multiple prediction algorithms.
