Molecular Methods

Cabana, Graciela S.; Hulsey, Brannon I; Pack, Frankie L.

doi:10.1016/b978-0-12-385189-5.00016-9

Cited by 1 publication

(1 citation statement)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Transposable elements are rare sequences of DNA that can move (or transpose) themselves to new positions within the genome of a single cell. A typical Alu element is an approximately 300 bp long transposable nucleotide sequence [6,7] found throughout the human genome [8]. The number of full-length or partial Alu elements in the human genome is estimated at over 1 million [9,10].…”

Section: Introductionmentioning

confidence: 99%

SPAST Intragenic CNVs Lead to Hereditary Spastic Paraplegia via a Haploinsufficiency Mechanism

Elert-Dobkowska,

Stepniak,

Radziwonik-Fraczyk

et al. 2024

IJMS

View full text Add to dashboard Cite

The most common form of hereditary spastic paraplegia (HSP), SPG4 is caused by single nucleotide variants and microrearrangements in the SPAST gene. The high percentage of multi-exonic deletions or duplications observed in SPG4 patients is predisposed by the presence of a high frequency of Alu sequences in the gene sequence. In the present study, we analyzed DNA and RNA samples collected from patients with different microrearrangements in SPAST to map gene breakpoints and evaluate the mutation mechanism. The study group consisted of 69 individuals, including 50 SPG4 patients and 19 healthy relatives from 18 families. Affected family members from 17 families carried varying ranges of microrearrangements in the SPAST gene, while one individual had a single nucleotide variant in the 5′UTR of SPAST. To detect the breakpoints of the SPAST gene, long-range PCR followed by sequencing was performed. The breakpoint sequence was detected for five different intragenic SPAST deletions and one duplication, revealing Alu-mediated microhomology at breakpoint junctions resulting from non-allelic homologous recombination in these patients. Furthermore, SPAST gene expression analysis was performed using patient RNA samples extracted from whole blood. Quantitative real-time PCR tests performed in 14 patients suggest no expression of transcripts with microrearrangements in 5 of them. The obtained data indicate that nonsense-mediated decay degradation is not the only mechanism of hereditary spastic paraplegia in patients with SPAST microrearrangements.

show abstract