Genetic basis of variable exon 9 skipping in cystic fibrosis transmembrane conductance regulator mRNA

Chu, Chin-Shyan; Trapnell, Bruce C.; Curristin, Sheila M.; Cutting, Garry R.; Crystal, Ronald G.

doi:10.1038/ng0293-151

Cited by 461 publications

(341 citation statements)

References 49 publications

Supporting

Mentioning

323

Contrasting

Unclassified

Order By: Relevance

“…23 This nonfunctional CFTR mRNA accounts for up to 92% of the total mRNA when the 5T allele is found on both CFTR genes. 11 We observed a significant proportion of Chinese CBAVD males who have the 5T allele (44.50%), as compared with the control males (13.46%). Also, a high frequency of homozygous (5T/5T) was found in the Chinese CBAVD patients ( Table 1).…”

Section: Cftr Polymorphisms In Chinese Males With Cbavd Wh Ni Et Al 688mentioning

confidence: 63%

“…10 The 7T or 9T allele generate a predominantly normal mRNA transcript, whereas the 5T variant affects splicing efficiency and results in reduced levels of normal mRNA due to deletion of exon 9. 11 The protein product of the CFTR transcript lacking exon 9 is devoid of cyclic adenosine monophosphate-activated chloride conductance, and therefore, the 5T allele is now considered as a mild mutation with an incomplete penetrance. 11,12 Prior studies have demonstrated that the disease penetrance of 5T depends on the copy number of its adjacent TG repeats.…”

Section: Introductionmentioning

confidence: 99%

“…11 The protein product of the CFTR transcript lacking exon 9 is devoid of cyclic adenosine monophosphate-activated chloride conductance, and therefore, the 5T allele is now considered as a mild mutation with an incomplete penetrance. 11,12 Prior studies have demonstrated that the disease penetrance of 5T depends on the copy number of its adjacent TG repeats. 13,14 The number of TG repeats immediately adjacent to 5T is associated with a variable efficiency of exon 9 splicing.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The CFTR polymorphisms poly-T, TG-repeats and M470V in Chinese males with congenital bilateral absence of the vas deferens

Ni¹,

Jiang²,

Fei³

et al. 2012

Asian J Androl

View full text Add to dashboard Cite

Congenital bilateral absence of the vas deferens (CBAVD) is a frequent cause of obstructive azoospermia, and mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene have also been frequently identified in patients with CBAVD. However, the distribution of the CFTR polymorphisms M470V, poly-T, TG-repeats and F508del mutation in the Chinese CBAVD population with presumed low cystic fibrosis (CF) frequency remains to be evaluated. Samples obtained from 109 Chinese infertile males with CBAVD and 104 normal controls were analyzed for the presence of CFTR (TG)m(T)n, M470V and F508del by PCR amplification followed by direct sequencing. Our study showed that the F508del mutation was not found in our patients. The 5T mutation was present with high frequency in Chinese CBAVD patients and IVS8-5T linked to either 12 or 13 TG repeats was highly prevalent among CBAVD patients (97.22% of 72 cases and 96.91% of 97 alleles with IVS8-5T). Moreover, a statistically significant relationship between TG12-5T-V470 haplotype and CBAVD was detected. This study indicated that the CFTR polymorphisms poly-T, TG-repeats and M470V might affect the process of CBAVD in the Chinese population.

show abstract

Section: Cftr Polymorphisms In Chinese Males With Cbavd Wh Ni Et Al 688mentioning

confidence: 63%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The CFTR polymorphisms poly-T, TG-repeats and M470V in Chinese males with congenital bilateral absence of the vas deferens

Ni¹,

Jiang²,

Fei³

et al. 2012

Asian J Androl

View full text Add to dashboard Cite

show abstract

“…16 The IVS-8 polymorphism affects the splicing efficiency of intron 8. 17,18 A tract of 7T or 9T at the 3Ј end of intron 8 insures proper splicing of the intron while a 5T results in a majority of mRNA lacking exon 9. 19 -22 Each mutation is independently considered mild because in both cases, residual activity of the ion channel remains.…”

mentioning

confidence: 99%

Long-Range (17.7 kb) Allele-Specific Polymerase Chain Reaction Method for Direct Haplotyping of R117H and IVS-8 Mutations of the Cystic Fibrosis Transmembrane Regulator Gene

Pont-Kingdon

Jama

Miller

et al. 2004

The Journal of Molecular Diagnostics

View full text Add to dashboard Cite

Genotyping of genetic polymorphisms is widely used in clinical molecular laboratories to confirm or predict diseases due to single locus mutations. In contrast, very few molecular methods determine the phase or haplotype of two or more mutations that are kilobases apart. In this report, we describe a new method for haplotyping based on long-range allelespecific PCR. Reaction conditions were established to circumvent the incompatibility of using allele-specific primers and a polymerase with proofreading activity. Detection of locus single mutations affecting gene function has been very successful in providing molecular diagnostics for human genetic diseases. In contrast, detection of two or more mutations on the same gene and determination of their phase (or haplotype) is more challenging. Advances in the field of human genome mapping, 1 the search for complex disease determinants, 2 pharmacogenomics, 3 and accumulation of data from mutation screening programs 4 underline the need to develop additional molecular haplotyping methods. Current technologies require physical separation of chromosomes or cloning or are limited to analysis of single nucleotide polymorphism (SNPs) present in the range of 1 kilobase (kb). [5][6][7][8][9][10][11][12][13][14] One recent exception is the development of a long-range molecular technique using a twostep PCR/ligation procedure. 15 Here we describe an assay that uses long-range allele-specific amplification to haplotype two mutations separated by 17.7 kb in the cystic fibrosis transmembrane regulator (CFTR) gene.The CFTR gene encodes a chloride channel and mutations in this gene are responsible for classic cystic fibrosis (CF) and atypical forms of the disease. Two of these mutations, R117H in exon 4 and the 5T polymorphism of the polythymidine tract in intron 8 (IVS-8 5T polymorphism) have a phenotypic synergic effect. The mutation R117H which accounts for approximately 0.8% of mutant alleles, changes an arginine to an histidine in a transmembrane domain of the protein, altering the conductance of the ion channel. 16 The IVS-8 polymorphism affects the splicing efficiency of intron 8. 17,18 A tract of 7T or 9T at the 3Ј end of intron 8 insures proper splicing of the intron while a 5T results in a majority of mRNA lacking exon 9. 19 -22 Each mutation is independently considered mild because in both cases, residual activity of the ion channel remains. Therefore, an individual carrying the R117H mutation and the IVS-8 5T polymorphism on two different chromosomes (in trans) is not affected by, nor considered a carrier of classic CF 23 although this individual might present with atypical CF. In contrast, a gene with both R117H and IVS-8 5T (in cis) is severely affected. 24 Both R117H and the IVS-8 5T variants have been found at higher frequencies in individuals with atypical CF than in the normal population. 22,[25][26][27] If an individual is heterozygous for both R117H and the IVS-8 5T variant, it is necessary to establish if both mutations are in cis or in trans to correctly analy...

show abstract

“…The probability of alternative splicing of exon 9 is related to the genotype of poly T and TG repeats in intron 8. 20 The analysis of transcripts is not routinely performed to detect the CFTR mutations in medical genetic laboratories, but a recent study identified a novel insertion between exons 10 and 11. 21 Another study of CF patients with only one mutation in the coding region of CFTR demonstrated a pathological reduction in the amount of RNA from the other allele in 5 of 11 patients.…”

Section: Discussionmentioning

confidence: 99%

Detection of a large heterozygous deletion and a splicing defect in the CFTR transcripts from nasal swab of a Japanese case of cystic fibrosis

et al. 2012

View full text Add to dashboard Cite

Genetic basis of variable exon 9 skipping in cystic fibrosis transmembrane conductance regulator mRNA

Cited by 461 publications

References 49 publications

The CFTR polymorphisms poly-T, TG-repeats and M470V in Chinese males with congenital bilateral absence of the vas deferens

The CFTR polymorphisms poly-T, TG-repeats and M470V in Chinese males with congenital bilateral absence of the vas deferens

Long-Range (17.7 kb) Allele-Specific Polymerase Chain Reaction Method for Direct Haplotyping of R117H and IVS-8 Mutations of the Cystic Fibrosis Transmembrane Regulator Gene

Detection of a large heterozygous deletion and a splicing defect in the CFTR transcripts from nasal swab of a Japanese case of cystic fibrosis

Contact Info

Product

Resources

About