title: Correction of Hair Shaft Defects through Allele-Specific Silencing of Mutant Krt75.
abstract: Dominant mutations in keratin genes can cause a number of inheritable skin disorders characterized by intraepidermal blistering, epidermal hyperkeratosis, or abnormalities in skin appendages, such as nail plate dystrophy and structural defects in hair. Allele-specific silencing of mutant keratins through RNA interference is a promising therapeutic approach for suppressing the expression of mutant keratins and related phenotypes in the epidermis. However, its effectiveness on skin appendages remains to be confirmed in vivo. In this study, we developed allele-specific small interfering RNAs capable of selectively suppressing the expression of a mutant Krt75, which causes hair shaft structural defects characterized by the development of blebs along the hair shaft in mice. Hair regenerated from epidermal keratinocyte progenitor cells isolated from mutant Krt75 mouse models reproduced the blebbing phenotype when grafted in vivo. In contrast, mutant cells manipulated with a lentiviral vector expressing mutant Krt75-specific short hairpin RNA (shRNA) persistently suppressed this phenotype. The phenotypic correction was associated with a significant reduction of mutant Krt75 mRNA in the skin grafts. Thus, data obtained from this study demonstrated the feasibility of utilizing RNA interference to achieve durable correction of hair structural phenotypes through allele-specific silencing of mutant keratin genes.
authors.affiliation: Institute of Laboratory Animal Science, Chinese Academy of Medical Science and Comparative Medical Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Beijing, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Pathology, Stony Brook University, Stony Brook, New York, USA; Graduate Program in Genetics, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Snedecor
authors.affiliation: Institute of Laboratory Animal Science, Chinese Academy of Medical Science and Comparative Medical Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Beijing, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Institute of Laboratory Animal Science, Chinese Academy of Medical Science and Comparative Medical Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Beijing, China.
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Institute of Laboratory Animal Science, Chinese Academy of Medical Science and Comparative Medical Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Beijing, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Dermatology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Evan C
authors.initials: EC
authors.lastname: Jones
authors.affiliation: Institute of Laboratory Animal Science, Chinese Academy of Medical Science and Comparative Medical Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Beijing, China.
authors.forename: Lianfeng
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Dermatology, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Clark
authors.affiliation: Department of Dermatology and Charles C. Gates Center for Regenerative Medicine and Stem Cell Biology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.
authors.forename: Dennis R
authors.initials: DR
authors.lastname: Roop
authors.affiliation: Institute of Laboratory Animal Science, Chinese Academy of Medical Science and Comparative Medical Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Beijing, China. Electronic address: qinchuan@pumc.edu.cn.
authors.forename: Chuan
authors.initials: C
authors.lastname: Qin
authors.affiliation: Institute of Laboratory Animal Science, Chinese Academy of Medical Science and Comparative Medical Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, Ministry of Health, Beijing, China; Department of Pathology, Stony Brook University, Stony Brook, New York, USA; Department of Dermatology, Stony Brook University, Stony Brook, New York, USA; Department of Dermatology and Charles C. Gates Center for Regenerative Medicine and Stem Cell Biology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. Electronic address: jiang.chen@stonybrook.edu.
authors.forename: Jiang
authors.initials: J
authors.lastname: Chen
mesh_terms: D000483:Alleles; D000818:Animals; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D020868:Gene Silencing; D005799:Genes, Dominant; D018859:Hair Follicle / Q000002:abnormalities* / Q000254:growth & development; D053553:Keratin-6 / Q000235:genetics*; D007633:Keratins / Q000235:genetics; D051379:Mice; D008817:Mice, Mutant Strains; D020125:Mutation, Missense; D011897:Random Allocation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D016038:Skin Transplantation / Q000379:methods
chemical_list: D053553:Keratin-6; C526574:Krt75 protein, mouse; D007633:Keratins