This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Al-Kateb, H. Articles by Bähring, S. Search for Related Content PubMed PubMed Citation Articles by Al-Kateb, H. Articles by Bähring, S. Pubmed/NCBI databases OMIM Substance via MeSH Genetics Home Reference

(Circulation Research. 2003;93:e49.)
© 2003 American Heart Association, Inc.

Letter to the Editor

A Splice Mutation in a Syrian Autosomal Recessive Hypercholesterolemia Family Causes a Two-Nucleotide Deletion of mRNA

Hussam Al-Kateb

Institute of Molecular Genetics, University of Heidelberg, Heidelberg, Germany, Franz Volhard Clinic, HELIOS Klinikum-Berlin, Medical Faculty of the Charité and, Max-Delbruck-Center for Molecular Medicine, Berlin, Germany, hussam-alkateb@usa.com

Ekkehard K.F. Bautz

Institute of Molecular Genetics, University of Heidelberg, Heidelberg, Germany

Friedrich C. Luft

Franz Volhard Clinic, HELIOS Klinikum-Berlin, Medical Faculty of the Charité and, Max-Delbruck-Center for Molecular Medicine, Berlin, Germany

Sylvia Bähring

Franz Volhard Clinic, HELIOS Klinikum-Berlin, Medical Faculty of the Charité and, Max-Delbruck-Center for Molecular Medicine, Berlin, Germany

An extract of the first 250 words of the full text is provided, because this article has no abstract.

To the Editor:

Autosomal recessive hypercholesterolemia (ARH) is a rare Mendelian dyslipidemia characterized by markedly elevated plasma low-density lipoprotein (LDL) cholesterol levels. ARH is caused by mutations in a protein that contains a phosphotyrosine binding domain (PTB) required for internalization of LDL in the liver.^1,2 LDL receptor function is defective in liver and lymphocytes but not in fibroblasts from ARH patients. We reported a Syrian ARH family earlier in Circulation Research and identified a mutation in the splice-acceptor site of intron 1 that converts the guanine of AG into cytosine AC.³ Splice mutations may cause cryptic site activation, intron retention, creation of new splice sites, or exon skipping.⁴ However, we had no access to ARH gene-expressing tissue. We therefore constructed ARH gene fragments from an affected and a nonaffected individual and transfected the fragments into hepatocellular carcinoma cells (HEPG2).

DNA fragments containing the 3' part of intron 1, exon 2, the entire intron 2, exon 3, and the 5' part of intron 3 were PCR-amplified from nonaffected and affected probands. Both fragments were subcloned into TOPO-XL vector (Invitrogen). Fragments containing the 5' noncoding part of exon 1, exon 1, and the 5' part of intron 1 were also amplified and were subcloned upstream to the previously cloned fragment. The final partial ARH gene, with and without mutation, was cloned into phCMV3/Xi expression vector (Gene Therapy Systems, San Diego, Calif). We did not include the entire intron 1 because we were concerned that the entire intron could be retained upon ARH . . . [Full Text of this Article]