McClive P, Pall G, Newton K, Lee M, Mullins J, Forrester L
Centre for Genome Research, University of Edinburgh, Scotland, United Kingdom.
Dev Dyn. 1998 Jun;212(2):267-76. doi: 10.1002/(SICI)1097-0177(199806)212:2<267::AID-AJA11>3.0.CO;2-1.
We describe the characterisation of three gene trap integrations in embryonic stem cells in which the lacZ reporter gene is repressed by retinoic acid (RA) in vitro and is expressed in the developing heart in vivo. In one of these, the gene trap vector has integrated into a gene that is located on chromosome 17 and is homologous to the human transcription factor gene, TFEB. Embryonic and adult cardiac expression of both the fusion transcript and the endogenous gene was confirmed. However, we show that the integration has not resulted in a null allele, because wild type transcripts, possibly resulting from splicing around the vector, are observed in homozygous tissue. The other two cardiac-expressing gene trap integrations have occurred into exons on chromosomes 1 and 5 and have used cryptic donor sites within the vector to generate functional fusion transcripts. One of these exon integrations results in a lethal neonatal phenotype.
我们描述了胚胎干细胞中三个基因陷阱整合的特征，其中lacZ报告基因在体外被视黄酸（RA）抑制，而在体内发育中的心脏中表达。在其中一个整合中，基因陷阱载体已整合到位于17号染色体上且与人转录因子基因TFEB同源的一个基因中。融合转录本和内源基因在胚胎期和成年期心脏中的表达均得到证实。然而，我们发现这种整合并未导致无效等位基因，因为在纯合组织中观察到了可能由载体周围剪接产生的野生型转录本。另外两个在心脏中表达的基因陷阱整合发生在1号和5号染色体的外显子中，并利用载体中的隐蔽供体位点产生功能性融合转录本。其中一个外显子整合导致致死性新生表型。
