PubMed Health⌕ Search

PubMed · 12900550

Ectopic XIST transcripts in human somatic cells show variable expression and localization.

Abstract

XIST encodes a functional RNA that is expressed exclusively from the inactive X in female mammals and is required for the silencing of most of the genes on the chromosome. XIST transcripts remain in the nucleus, and their specific localization to the inactive X is important for silencing; however, it is not known how these transcripts localize to the inactive X chromosome. Expression of mouse and human XIST from ectopic sites has suggested that localization to the chromosome from which the gene is expressed may be dependent upon either the copy number of the integrated constructs or the level of ectopic XIST expression. To further examine the behavior of XIST transgenes when expressed from ectopic sites, we introduced an XIST-containing PAC into the human male somatic cell line HT-1080. In five different transformant clones, the degree of localization and associated DNA condensation of the surrounding chromatin varied within nuclei of the same clone, as well as among different clones. Comparing the number of integrated transgenes and the levels of XIST expression revealed that neither factor was sufficient for a tight localization of the XIST signal. Therefore, the extent of expression and localization of XIST transcripts from ectopic transgenes is likely dependent upon many interacting factors, including the number of integrated transgenes, the level of XIST expression, and the site of integration.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

J C Chow, L L Hall, J B Lawrence, C J Brown. 2002. Ectopic XIST transcripts in human somatic cells show variable expression and localization.. https://doi.org/10.1159/000071579

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

Protocol to improve isoform-level quantification of low-abundance transcripts via STALARD pre-amplification.

STALARD (selective target amplification for low-abundance RNA detection) enables isoform-level quantification of low-abundance RNAs using conventional laboratory equipment. Here, we describe steps for RNA isolation, primer design, reverse transcription, selective target amplification, and downstream analysis. The protocol couples selective pre-amplification with a quantitative reverse-transcription PCR (RT-qPCR) readout and optional nanopore sequencing. Using 1 μg input RNA and 12 pre-amplification cycles, STALARD reduces Cq values by approximately 10-12 cycles, bringing the target into a reliably quantifiable range. For complete details on the use and execution of this protocol, please refer to Jeong et al.1.

Gene Expression↗

Measures of genotype versus gene products: promise and pitfalls in cancer prevention.

We present a critical assessment of the promise and pitfalls of genotype versus gene function measures in cancer epidemiology studies. While both measures have pros and cons and are complementary, in terms of potential for contributing knowledge that directly leads to prevention, we argue that attention should be given to research relating functional parameters (single protein expression, functional or phenotypic assays or patterns of gene/protein expression) to disease risk. We present the theoretical and conceptual basis of why studies focusing on polymorphisms in the low-penetrance genes may be logically less fruitful for making inroads to cancer prevention than appropriately designed studies using validated functional parameters. We then substantiate these arguments with some concrete examples based on the current literature. We also discuss the limitations of including functional parameters in epidemiological studies and technical improvements required before such studies can truly fulfil their promise. Finally, we offer some specific recommendations for future research directions in this area.

Gene Expression↗

Specificity of short interfering RNA determined through gene expression signatures.

Short interfering RNA (siRNA) is widely used for studying gene function and holds great promise as a tool for validating drug targets and treating disease. A critical assumption in these applications is that the effect of siRNA on cells is specific, i.e., limited to the specific knockdown of the target gene. In this article, we characterize the specificity of siRNA by applying gene expression profiling. Several siRNAs were designed against different regions of the same target gene for three different targets. Their effects on cells were compared by using DNA microarrays to generate gene expression signatures. When the siRNA design and transfection conditions were optimized, the signatures for different siRNAs against the same target were shown to correlate very closely, whereas the signatures for different genes revealed no correlation. These results indicate that siRNA is a highly specific tool for targeted gene knockdown, establishing siRNA-mediated gene silencing as a reliable approach for large-scale screening of gene function and drug target validation.

Gene Expression↗