Lu Zhi Hong, Books Jason T, Kaufman Richard M, Ley Timothy J
Section of Stem Cell Biology, Division of Oncology, Washington University School of Medicine, St Louis, MO 63110, USA.
Blood. 2003 Aug 15;102(4):1531-3. doi: 10.1182/blood-2003-03-0708. Epub 2003 May 1.
The correction of mutant beta-globin genes has long been a therapeutic goal for patients with beta-thalassemia or hemoglobinopathies. The use of homologous recombination (HR) to achieve this goal is an attractive approach because it eliminates the need to include regulatory sequences in the therapeutic construct, and it eliminates mutagenesis induced by random integration. However, HR is a very inefficient process for gene correction, and its efficiency is probably locus dependent. The length of targeting arms is thought to be a determinant of targeting efficiency, so we compared the ability of standard (8-kb) versus very long (16-, 24-, and 110-kb) regions of homology to correct a mutant murine beta-globin gene in embryonic stem cells. Increasing the length of the targeting sequences did not increase the efficiency of HR in this locus, suggesting that alternative approaches will be required to improve the efficiency of this approach for globin gene correction.
长期以来，纠正突变的β-珠蛋白基因一直是β地中海贫血或血红蛋白病患者的治疗目标。利用同源重组（HR）来实现这一目标是一种有吸引力的方法，因为它无需在治疗构建体中包含调控序列，并且消除了随机整合诱导的诱变。然而，HR用于基因校正的过程效率非常低，其效率可能取决于基因座。靶向臂的长度被认为是靶向效率的一个决定因素，因此我们比较了标准（8kb）与非常长（16kb、24kb和110kb）同源区域在胚胎干细胞中校正突变小鼠β-珠蛋白基因的能力。增加靶向序列的长度并未提高该基因座处HR的效率，这表明需要采用其他方法来提高这种用于珠蛋白基因校正方法的效率。
