Activating the synthesis of progerin, the mutant prelamin A in Hutchinson-Gilford progeria syndrome, with antisense oligonucleotides.
Activating the synthesis of progerin, the mutant prelamin A in Hutchinson-Gilford progeria syndrome, with antisense oligonucleotides.
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DOI：
10.1093/hmg/ddp184
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发表时间：
2009-07-01
影响因子：
3.5
通讯作者：
Young SG
中科院分区：
文献类型：
--
作者：
Fong LG;Vickers TA;Farber EA;Choi C;Yun UJ;Hu Y;Yang SH;Coffinier C;Lee R;Yin L;Davies BS;Andres DA;Spielmann HP;Bennett CF;Young SG
Hutchinson-Gilford早衰综合征（HGPS）是由LMNA（编码层蛋白C和前层蛋白A的基因）外显子11上一个替代剪接供体位点的利用增加的点突变引起的。交替剪接减少了野生型prelamin A的转录本，增加了截断的prelamin A （progerin）的转录本。本研究表明，在野生型和HGPS成纤维细胞中，位于11外显子剪接供体位点下游的11外显子序列上的反义寡核苷酸（ASOs）促进了剪接的交替，增加了progerin的合成。事实上，转染这些ASOs的野生型成纤维细胞显示出与HGPS患者成纤维细胞相似（或更高）的早孕蛋白水平。根据代谢标记研究，这种蛋白被法酰化了。在野生型成纤维细胞中，progerin的合成伴随着与HGPS成纤维细胞相同的核形状和基因表达扰动。内含子11的5 '部分对应的ASO也促进了交替剪接。相比之下，针对外显子11序列5 '到交替剪接位点的ASO减少了HGPS细胞的交替剪接，并适度降低了progerin水平。因此，不同的ASOs可以用来增加或减少“HGPS拼接”。ASOs是在野生型细胞中重建HGPS病理生理的一种新的有力工具。
Hutchinson–Gilford progeria syndrome (HGPS) is caused by point mutations that increase utilization of an alternate splice donor site in exon 11 of LMNA (the gene encoding lamin C and prelamin A). The alternate splicing reduces transcripts for wild-type prelamin A and increases transcripts for a truncated prelamin A (progerin). Here, we show that antisense oligonucleotides (ASOs) against exon 11 sequences downstream from the exon 11 splice donor site promote alternate splicing in both wild-type and HGPS fibroblasts, increasing the synthesis of progerin. Indeed, wild-type fibroblasts transfected with these ASOs exhibit progerin levels similar to (or greater than) those in fibroblasts from HGPS patients. This progerin was farnesylated, as judged by metabolic labeling studies. The synthesis of progerin in wild-type fibroblasts was accompanied by the same nuclear shape and gene-expression perturbations observed in HGPS fibroblasts. An ASO corresponding to the 5′ portion of intron 11 also promoted alternate splicing. In contrast, an ASO against exon 11 sequences 5′ to the alternate splice site reduced alternate splicing in HGPS cells and modestly lowered progerin levels. Thus, different ASOs can be used to increase or decrease ‘HGPS splicing’. ASOs represent a new and powerful tool for recreating HGPS pathophysiology in wild-type cells.
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DOI：
10.1073/pnas.0505767102
发表时间：
2005-09-06
影响因子：
11.1
作者：
Toth, JI;Yang, SH;Fong, LG
通讯作者：
Fong, LG
DOI：
10.1073/pnas.0506001102
发表时间：
2005-09-06
影响因子：
11.1
作者：
Capell, BC;Erdos, MR;Collins, FS
通讯作者：
Collins, FS
影响因子：
82.9
作者：
Scaffidi, P;Misteli, T
通讯作者：
Misteli, T
影响因子：
21.3
作者：
Scaffidi, Paola;Misteli, Tom
通讯作者：
Misteli, Tom
影响因子：
9.8
作者：
Hua, Yimin;Vickers, Timothy A;Baker, Brenda F;Bennett, C Frank;Krainer, Adrian R
通讯作者：
Krainer, Adrian R