FES-Cre targets phosphatidylinositol glycan class A (PIGA) inactivation to hematopoietic stem cells in the bone marrow.
FES-Cre targets phosphatidylinositol glycan class A (PIGA) inactivation to hematopoietic stem cells in the bone marrow.
复制标题
DOI：
10.1084/jem.194.5.581
复制
发表时间：
2001-09-03
期刊：
影响因子：
--
通讯作者：
Bessler M
中科院分区：
文献类型：
--
作者：
Keller P;Payne JL;Tremml G;Greer PA;Gaboli M;Pandolfi PP;Bessler M
X-连锁磷脂酰肌醇聚糖A类（PIGA）基因的体细胞突变导致阵发性睡眠性血红蛋白尿症患者血细胞上糖基磷脂酰肌醇（GPI）连接蛋白的丢失。因为所有的血细胞谱系都可能受到影响，所以认为突变发生在造血干细胞中。在转基因小鼠中，非活性Piga基因的生殖系传播是胚胎致死的。为了在早期造血中抑制小鼠Piga基因，我们因此选择使用Cre/loxP系统的条件性基因失活。我们在人c-fes基因的转录调控序列下表达Cre重组酶。FES-Cre使携带floxed Piga等位基因的小鼠（LF小鼠）的造血细胞中的PIGA失活。PIGA−细胞存在于所有定型造血谱系中，但不存在于原始造血。它们在新生小鼠中的比例很低，但随后持续增加，首次产生几乎完全具有PIGA-血细胞的小鼠。GPI连接蛋白的丢失主要发生在CFU-GEMM阶段之前的c-kit+ CD 34 +Lin−祖细胞中。使用纯化的PIGA−细胞进行骨髓重建实验，我们证明LF小鼠具有缺乏GPI连接蛋白的长期骨髓重建细胞，这表明在造血干细胞中发生了floxed Piga等位基因的重组。
A somatic mutation in the X-linked phosphatidylinositol glycan class A (PIGA) gene causes the loss of glycosyl phosphatidylinositol (GPI)-linked proteins on blood cells from patients with paroxysmal nocturnal hemoglobinuria. Because all blood cell lineages may be affected it is thought that the mutation occurs in a hematopoietic stem cell. In transgenic mice, germline transmission of an inactive Piga gene is embryonic lethal. To inactivate the murine Piga gene in early hematopoiesis we therefore chose conditional gene inactivation using the Cre/loxP system. We expressed Cre recombinase under the transcription regulatory sequences of the human c-fes gene. FES-Cre inactivated PIGA in hematopoietic cells of mice carrying a floxed Piga allele (LF mice). PIGA− cells were found in all hematopoietic lineages of definitive but not primitive hematopoiesis. Their proportions were low in newborn mice but subsequently increased continuously to produce for the first time mice that have almost exclusively PIGA− blood cells. The loss of GPI-linked proteins occurred mainly in c-kit+CD34+Lin− progenitor cells before the CFU-GEMM stage. Using bone marrow reconstitution experiments with purified PIGA− cells we demonstrate that LF mice have long-term bone marrow repopulating cells that lack GPI-linked proteins, indicating that recombination of the floxed Piga allele occurs in the hematopoietic stem cell.
登录
查看更多内容
影响因子：
20.3
作者：
Kawagoe, K;Kitamura, D;Takeda, J
通讯作者：
Takeda, J
DOI：
10.1073/pnas.82.8.2379
发表时间：
1985-01-01
影响因子：
11.1
作者：
FELDMAN, RA;GABRILOVE, JL;HANAFUSA, H
通讯作者：
HANAFUSA, H
影响因子：
5.3
作者：
GREER, P;HAIGH, J;PAWSON, T
通讯作者：
PAWSON, T
影响因子：
20.3
作者：
Fisher, RC;Lovelock, JD;Scott, EW
通讯作者：
Scott, EW
影响因子：
64.5
作者：
ELLIS, L;CLAUSER, E;RUTTER, WJ
通讯作者：
RUTTER, WJ