PCR amplification of chromosome-specific alpha satellite DNA: Definition of centromeric STS markers and polymorphic analysis

Warburton, Peter E.; Greig, Gillian; Haaf, Thomas; Willard, Huntington F.

doi:10.1016/0888-7543(91)90139-6

Cited by 103 publications

(84 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A 850 bp fragment was amplified with primers which were varied according to Warburton et al (1991). A BLAST-search of this human sequence fragment in the EMBL rodent DNA database as well as in the ENSEMBLE whole mouse genome browser (EBI, Hinxton, Cambridge, UK) revealed no significant similarity with any mouse DNA sequence.…”

Section: Discussionmentioning

confidence: 99%

“…The presence of human-specific DNA within the blood and organs of transplanted mice was confirmed by polymerase chain reaction (PCR) amplifying an 850-bp fragment of the a-satellite region of the human chromosome 17 using primers corresponding to the primer pair 17a1/17a2 as described by Warburton et al (1991). The primers were elongated to 25 nucleotides each for use at higher annealing temperatures.…”

Section: Conventional Dna Pcrmentioning

confidence: 99%

“…Therefore, we decided to develop a sensitive PCR assay which is universally applicable either for the detection of human cells in murine blood and organs or for a quantification of such cells via real-time (TaqMan) PCR. We targeted for that purpose a highly repetitive a-satellite DNA sequence of the centromer region of human chromosome 17 with primers which were specifically modified by us according to Warburton et al (1991). The sensitivity and specificity of that method was investigated and used for: (i) the detection of micrometastases of a human breast carcinoma xenograft; (ii) the estimation of dissemination of a human acute lymphatic leukaemia (ALL) model; (iii) the proof of distribution of mature lymphocytes and (iv) the evaluation of the engraftment potential of human blood stem cells.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems

et al. 2002

View full text Add to dashboard Cite

The sensitive detection of human cells in immunodeficient rodents is a prerequisite for the monitoring of micrometastasis of solid tumours, dissemination of leukaemic cells, or engraftment of haematological cells. We developed a universally applicable polymerase chain reaction method for the detection of a human-specific 850-bp fragment of the a-satellite DNA on human chromosome 17. The method allows the detection of one human cell in 10 6 murine cells and could be established as both, a conventional DNA polymerase chain reaction-assay for routine screening, and a quantitative real-time polymerase chain reaction-assay using TaqMan-methodology. It was applied to the following xenotransplantation systems in SCID and NOD/ SCID mice: (1) In a limiting dilution assay, cells of the MDA-MB 435 breast carcinoma were injected into the mammary fat pad of NOD/SCID mice. It could be shown that 10 cells mouse 71 were sufficient to induce a positive polymerase chain reaction signal in liver and lung tissue 30 days after transplantation as an indicator for micrometastasis. At this time a palpable tumour was not yet detectable in the mammary fat pad region. (2) Cells of a newly established human acute lymphatic leukaemia were administered intraperitoneally to SCID mice. These cells apparently disseminated and were detectable as early as day 50 in the peripheral blood of living mice, while the leukaemia manifestation was delayed by day 140. (3) In a transplantation experiment using mature human lymphocytes we wanted to standardise conditions for a successful survival of these cells in NOD/SCID mice. It was established that at least 5610 7 cells given intravenously were necessary and that the mice had to be conditioned by 2 Gy body irradiation to get positive polymerase chain reaction bands in several organs. (4) Engraftment studies with blood stem cells originating from cytapheresis samples of tumour patients or from cord blood were undertaken in NOD/SCID mice in order to define conditions of successful engraftment and to use this model for further optimisation strategies. The polymerase chain reaction method presented allowed a reliable prediction of positive engraftment and agreed well with the results of immunohistochemical or FACS analysis. All together, the polymerase chain reaction method developed allows a sensitive and reliable detection of low numbers of human cells in immunodeficient hosts. In combination with real-time (TaqMan) technique it allows an exact quantification of human cells. As this method can be performed with accessible material of living animals, follow up studies for the monitoring of therapeutic interventions are possible in which the survival time of mice as evaluation criteria can be omitted.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Conventional Dna Pcrmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems

et al. 2002

View full text Add to dashboard Cite

show abstract

“…The copy number of the X chromosome was examined using specific alpha satellite DNA generated by PCR. (23) Expression analysis of WTX mRNA. Total RNA was extracted from 49 WTs and four normal kidney tissues adjacent to WT or congenital mesoblastic nephroma.…”

Section: Methodsmentioning

confidence: 99%

Different incidences of epigenetic but not genetic abnormalities between Wilms tumors in Japanese and Caucasian children

et al. 2012

View full text Add to dashboard Cite

Epidemiological studies show that the incidence of Wilms tumor (WT) in East-Asian children is half of that in Caucasian children. Abnormalities of WT1, CTNNB1, WTX, and IGF2 were reported to be involved in Wilms tumorigenesis in Caucasians, although none of the studies simultaneously evaluated the four genes. WTX forms the b-catenin degradation complex; however, the relationship between WTX abnormality and CTNNB1 mutation was uncertain in WTs. We examined abnormalities of the four genes in 114 Japanese with WTs to clarify the relationship between genetic and epigenetic factors and the incidence of WTs. We found that abnormalities of WTX and CTNNB1 were mutually exclusive, and that although CTNNB1 mutation was frequent in WTs with WT1 abnormality, but rare in WTs without, the incidences of WTX abnormality were similar between WTs with or without WT1 abnormality. These findings were consistent with those reported in Caucasian populations, and indicate multiple roles of WTX abnormality. Abnormalities of WT1, WTX and CTNNB1, and loss of IGF2 imprinting (LOI) were detected in 31.6%, 22.8%, 26.3%, and 21.1% of the 114 WTs, respectively. When we selected 101 sporadic WTs, the incidences of WT1, CTNNB1, or WTX abnormality were generally comparable between the two populations, whereas the incidence of IGF2 LOI was lower in Japanese than that of IGF2 LOI reported in Caucasians (P = 0.04). This is the first comprehensive study of the four genes, and the results supported the hypothesis that the lower incidence of IGF2 LOI contributes to the lower incidence of WTs in Japanese children. (Cancer Sci 2012; 103: 1129-1135 T he rates of various types of adult cancers, including prostate, lung, breast, liver and gastric cancers, differ among ethnic populations, which are thought to be caused by different environmental factors. (1) In contrast, most pediatric tumors such as neuroblastomas and retinoblastomas show similar rates among ethnic populations. An exception is Wilms tumor (WT), a common pediatric tumor of the kidney, which is known to have different incidence rates between East-Asian and Caucasian populations. (2) Wilms tumor accounts for 8% of childhood cancers and occurs in 1 in 10 000 Caucasian children, though its incidence in East-Asian populations is half of that. (2) In Hawaii and Britain, the incidence of WT in people of Asian descent is about half to two-thirds of that in Caucasians, (3,4) suggesting that environmental factors play little part in the lower incidence.IGF2 is an imprinting gene expressed from the paternally inherited allele, and encodes a fetal polypeptide growth factor. Loss of imprinting (LOI) of IGF2 has been reported in various tumors, including pediatric tumors. There have been conflicting reports on the rates of IGF2 LOI in Japanese with WTs; one study reported IGF2 LOI in none of 21 tumors, whereas we previously reported it in seven of 27 tumors. (5,6) These results prompted us to analyze the IGF2 imprinting status in a substantial number of Japanese children with WTs.Various abnormalities of...

show abstract

“…Centromeric sequences vary considerably between individuals (Wevrick and Willard 1989;Warburton et al 1991), introducing new datasets to the study of human genetic diversity, evolution, and disease. As a consequence of this variation, each human genome is expected to contain a personalized inventory of centromere sequence composition and organization.…”

Section: Introductionmentioning

confidence: 99%

Human centromere genomics: now it's personal

Hayden

2012

Chromosome Res

View full text Add to dashboard Cite

Advances in human genomics have accelerated studies in evolution, disease, and cellular regulation. However, centromere sequences, defining the chromosomal interface with spindle microtubules, remain largely absent from ongoing genomic studies and disconnected from functional, genome-wide analyses. This disparity results from the challenge of predicting the linear order of multi-megabase-sized regions that are composed almost entirely of nearidentical satellite DNA. Acknowledging these challenges, the field of human centromere genomics possesses the potential to rapidly advance given the availability of individual, or personalized, genome projects matched with the promise of long-read sequencing technologies. Here I review the current genomic model of human centromeres in consideration of those studies involving functional datasets that examine the role of sequence in centromere identity.

show abstract

PCR amplification of chromosome-specific alpha satellite DNA: Definition of centromeric STS markers and polymorphic analysis

Cited by 103 publications

References 39 publications

Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems

Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems

Different incidences of epigenetic but not genetic abnormalities between Wilms tumors in Japanese and Caucasian children

Human centromere genomics: now it's personal

Contact Info

Product

Resources

About