Tags

Type your tag names separated by a space and hit enter

Disease-causing variants of the conserved +2T of 5' splice sites can be rescued by engineered U1snRNAs.
Hum Mutat. 2019 01; 40(1):48-52.HM

Abstract

The ability of variants of the spliceosomal U1snRNA to rescue splicing has been proven in several human disease models, but not for nucleotide changes at the conserved GT nucleotide of 5' splice sites (5'ss), frequent and associated with severe phenotypes. Here, we focused on variants at the 5'ss of F9 intron 3, leading to factor IX (FIX) deficiency (hemophilia B). Through minigene expression, we demonstrated that all changes induce complete exon 3 skipping, which explains the associated hemophilia B phenotype. Interestingly, engineered U1snRNAs remarkably increased the proportion of correct transcripts in the presence of the c.277+4A>G (∼60%) and also c.277+2T>C mutation (∼20%). Expression of splicing-competent cDNA constructs indicated that the splicing rescue produces an appreciable increase of secreted FIX protein levels. These data provide the first experimental evidence that even part of variants at the conserved 5'ss +2T nucleotide can be rescued, thus expanding the applicability of this U1snRNA-based approach.

Authors+Show Affiliations

Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.Department of Experimental and Diagnostic Medicine, University of Ferrara, Ferrara, Italy.Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30408273

Citation

Scalet, Daniela, et al. "Disease-causing Variants of the Conserved +2T of 5' Splice Sites Can Be Rescued By Engineered U1snRNAs." Human Mutation, vol. 40, no. 1, 2019, pp. 48-52.
Scalet D, Maestri I, Branchini A, et al. Disease-causing variants of the conserved +2T of 5' splice sites can be rescued by engineered U1snRNAs. Hum Mutat. 2019;40(1):48-52.
Scalet, D., Maestri, I., Branchini, A., Bernardi, F., Pinotti, M., & Balestra, D. (2019). Disease-causing variants of the conserved +2T of 5' splice sites can be rescued by engineered U1snRNAs. Human Mutation, 40(1), 48-52. https://doi.org/10.1002/humu.23680
Scalet D, et al. Disease-causing Variants of the Conserved +2T of 5' Splice Sites Can Be Rescued By Engineered U1snRNAs. Hum Mutat. 2019;40(1):48-52. PubMed PMID: 30408273.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Disease-causing variants of the conserved +2T of 5' splice sites can be rescued by engineered U1snRNAs. AU - Scalet,Daniela, AU - Maestri,Iva, AU - Branchini,Alessio, AU - Bernardi,Francesco, AU - Pinotti,Mirko, AU - Balestra,Dario, Y1 - 2018/11/19/ PY - 2018/09/26/received PY - 2018/10/26/revised PY - 2018/11/05/accepted PY - 2018/11/9/pubmed PY - 2020/3/7/medline PY - 2018/11/9/entrez KW - ExSpeU1 KW - RNA splicing KW - hemophilia B KW - human disease KW - splicing mutations SP - 48 EP - 52 JF - Human mutation JO - Hum. Mutat. VL - 40 IS - 1 N2 - The ability of variants of the spliceosomal U1snRNA to rescue splicing has been proven in several human disease models, but not for nucleotide changes at the conserved GT nucleotide of 5' splice sites (5'ss), frequent and associated with severe phenotypes. Here, we focused on variants at the 5'ss of F9 intron 3, leading to factor IX (FIX) deficiency (hemophilia B). Through minigene expression, we demonstrated that all changes induce complete exon 3 skipping, which explains the associated hemophilia B phenotype. Interestingly, engineered U1snRNAs remarkably increased the proportion of correct transcripts in the presence of the c.277+4A>G (∼60%) and also c.277+2T>C mutation (∼20%). Expression of splicing-competent cDNA constructs indicated that the splicing rescue produces an appreciable increase of secreted FIX protein levels. These data provide the first experimental evidence that even part of variants at the conserved 5'ss +2T nucleotide can be rescued, thus expanding the applicability of this U1snRNA-based approach. SN - 1098-1004 UR - https://www.unboundmedicine.com/medline/citation/30408273/Disease_causing_variants_of_the_conserved_+2T_of_5'_splice_sites_can_be_rescued_by_engineered_U1snRNAs_ DB - PRIME DP - Unbound Medicine ER -