Ectopic gene targeting exhibits a bimodal distribution of integration in murine cells, indicating that both intra- and interchromosomal sites are accessible to the targeting vector.

G Dellaire; N Lemieux; A Belmaaza; P Chartrand

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Ectopic gene targeting exhibits a bimodal distribution of integration in murine cells, indicating that both intra- and interchromosomal sites are accessible to the targeting vector.

机译：异位基因靶向在鼠细胞中表现出整合的双峰分布，表明靶向载体可访问染色体内和染色体间位点。

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Ectopic gene targeting is an alternative outcome of the gene targeting process in which the targeting vector acquires sequences from the genomic target but proceeds to integrate elsewhere in the genome. Using two-color fluorescent in situ hybridization analysis, we have determined the integration sites of the gene targeting vector with respect to the target locus in a murine fibroblast line (LTA). We found that for ectopic gene targeting the distribution of integration sites was bimodal, being either within 3 Mb of the target or on chromosomes distinct from the chromosome carrying the target locus. Inter- and intrachromosomal sites appeared to be equally accessible to the targeting vector, with site-specific variations. Interestingly, interphase analysis indicated that vector sequences which had integrated ectopically in chromosomes other than the target colocalized with the target locus at a significant frequency compared to that of colocalization to random unlinked loci. We propose that ectopic gene targeting could be used to determine which chromosomal domains within the genome are accessible to a given genetic locus. Thus, recombination access mapping may present a new paradigm for the analysis of DNA accessibility and interaction within the genome.

机译：异位基因靶向是基因靶向过程的另一种结果，其中靶向载体从基因组靶标获取序列，然后继续整合到基因组中的其他位置。使用双色荧光原位杂交分析，我们已经确定了基因靶向载体相对于鼠成纤维细胞系（LTA）中靶基因座的整合位点。我们发现，针对异位基因靶向整合位点的分布是双峰的，位于靶标的3 Mb内或位于不同于携带靶标基因座的染色体的染色体上。染色体间和染色体内位点似乎对靶向载体具有同等的访问权限，但具有位点特异性变异。有趣的是，相间分析表明，与共定位到随机未连锁基因座的频率相比，异位整合在非目标染色体上的载体序列与目标基因座共定位的频率很高。我们提议异位基因靶向可用于确定基因组内的哪些染色体结构域可访问给定的基因座。因此，重组访问图谱可为分析DNA可及性和基因组内相互作用提供新的范例。

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《Molecular and Cellular Biology》 |1997年第9期|共10页
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G Dellaire; N Lemieux; A Belmaaza; P Chartrand;
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6. Ectopic gene targeting exhibits a bimodal distribution of integration in murine cells indicating that both intra- and interchromosomal sites are accessible to the targeting vector. [O] . G Dellaire, N Lemieux, A Belmaaza, 1997

机译：异位基因靶向在鼠细胞中表现出整合的双峰分布表明靶向载体可访问染色体内和染色体间位点。
7. Ectopic gene targeting exhibits a bimodal distribution of integration in murine cells, indicating that both intra- and interchromosomal sites are accessible to the targeting vector. [O] . Dellaire, G, Lemieux, N, Belmaaza, A, 1997

机译：异位基因靶向在鼠细胞中表现出整合的双峰分布，表明靶向载体可访问染色体内和染色体间位点。

Ectopic gene targeting exhibits a bimodal distribution of integration in murine cells, indicating that both intra- and interchromosomal sites are accessible to the targeting vector.

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