CRISPR base editor treats premature-aging syndrome.
CRISPR base editor treats premature-aging syndrome.
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CRISPR基础编辑器治疗早衰综合征。
DOI：
10.1038/s41392-021-00576-6
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发表时间：
2021-04-16
影响因子：
39.3
通讯作者：
Wu M
中科院分区：
文献类型：
--
作者：
Lin P;Jiang J;Wu M
Koblan等人最近发表在Nature上的一篇论文报道了使用CRISPR介导的腺嘌呤碱基编辑器（ABE）修复Hutchinson-Gilford早衰综合征（HGPS或早衰症）的突变，减轻症状并延长小鼠的寿命（图1），1代表了人类加速衰老疾病和潜在其他遗传疾病治疗设计的重大进展。HGPS是一种极其罕见但无法治愈的遗传性疾病，表现出多个器官的快速老化，最终导致早期死亡（患者在十几岁或20岁出头时死亡）。大多数HGPS患者表现出核纤层蛋白A（LMNA）基因中从胞嘧啶（C）到胸腺嘧啶（T）的单碱基突变（1824 C> T）（图1a）。该突变导致LMNA转录期间的错误剪接，随后翻译成截短的核纤层蛋白A蛋白，称为早老蛋白（图1a），其中早老蛋白总是保持用法尼基基团标记。法尼基化早老蛋白的积累对核形状和刚性是有毒的，这阻碍了核功能，导致HGPS疾病。发现早老症治疗方法的尝试最初集中在试图减少法尼基化早老蛋白的积累。虽然已经筛选和测试了法尼基转移酶的小分子抑制剂，为患者提供治疗选择，但在临床上使用它们的治疗只能部分缓解疾病症状。Beyret等人和Mrsago-Fernández等人通过破坏HGPS突变基因的活性使用CRISPR-Cas9介导的核纤层蛋白A/早老蛋白减少。但他们的健康状况只恢复了HGPS细胞和小鼠的几处变化。更糟糕的是，这些治疗策略还导致LMNA基因的移码突变，这可能会使人类的基因拷贝无法充分纠正这种疾病。因此，有必要开发一种新的策略来直接修复导致HGPS的突变。CRISPR-Cas生物技术、基因修饰和工程正在通过纠正基因突变来彻底改变治疗遗传疾病的方式，这有助于基因编辑治疗或治愈某些遗传性疾病、癌症和其他疾病。ABE使用与进化的脱氧腺苷脱氨酶融合的催化受损的Cas9，在DNA复制期间通过聚合酶介导靶向A· T向G· C的转化，该中间体读作鸟嘌呤（G）。2 Koblan等人使用优化的ABE 7。10变体（ABEmax-VRQR）3，具有靶向sgRNA的c. LMNA中1824 C> T（图1 B）。1通过慢病毒递送，Koblan等人在来自HGPS患者的两个原代细胞中测试了ABEmax-VRQR，导致约90%的突变基因得到纠正。1这导致了核纤层蛋白A的正常剪接，减少了早老蛋白的显著表达和积累，并基本上纠正了异常的核（图1 B）。因此，本研究开发的ABE方法可以修复HGPS细胞中的突变基因。接下来，为了研究使用ABE治疗来纠正体内HGPS突变，Koblan等人使用临床腺相关病毒载体共包装ABEmax-VRQR和sgRNA（ABE-AAV），将修复复合物机制递送到HGPS小鼠模型中（图lc）1-C57 BL/6小鼠纯合包括致病性人LMNA c。1824 C> T等位基因，其发展在人类HGPS患者中观察到的许多标志性症状，例如血管平滑肌细胞（VSMC）损失、心血管并发症、皮下脂肪损失和早期死亡。4在出生后第3天（P3）和第14天（P14）的小鼠中眶后注射ABE-AAV后，Koblan等人实现了10-60%的人...
A recent paper published in Nature by Koblan et al. reported the use of CRISPR-mediated adenine base editor (ABE) to repair mutations of the Hutchinson–Gilford progeria syndrome (HGPS or progeria), attenuate symptoms, and extend lifespan of mice (Fig. 1), 1 representing a major advance in design of treatments for human accelerated-ageing disorders and potentially other genetic diseases. HGPS is an extremely rare but incurable genetic disease exhibiting rapid aging of multiple organs, ultimately leading to early death (patients die at teens or early 20s). Most HGPS patients exhibit a single base mutation in the lamin A (LMNA) gene from cytosine (C) to a thymine (T)(1824C> T)(Fig. 1 a). This mutation leads to mis-splicing during LMNA transcription and subsequently translates into a truncated lamin A protein, termed progerin (Fig. 1 a), in which progerin protein always keeps tagging with a farnesyl group. The accumulation of farnesylated progerin is toxic for nuclear shape and rigidity, which hampers nucleus function resulting in HGPS disease. Attempts to discover treatments for progeria initially focused on trying to reduce the accumulation of farnesylated progerin. Although small-molecule inhibitors for farnesyltransferase have been screened and tested to provide treatment options for patients, the treatment with them only partially alleviates the disease symptoms in clinics. Beyret et al. and Santiago-Fernández et al. have used CRISPR-Cas9-mediated lamin A/progerin reduction by disrupting activity of HGPS-mutated gene. But their health reverted only several alterations in HGPS cells and mice. To make the matter worse, these treatment strategies also lead to frameshift mutations in the LMNA gene, which could disable a human’s copy of the gene to adequately correct the disease due to this harmful effect. Therefore, it is necessary to develop a new strategy to directly repair the mutation that causes HGPS. CRISPR-Cas biotechnology, gene-modifying, and engineering are revolutionizing the ways of treating genetic diseases by correcting genetic mutations, which facilitate gene editing to treat or cure certain inheritable diseases, cancers, and other illnesses. ABE, using a catalytically impaired Cas9 fused with evolved deoxyadenosine deaminase, mediates the conversion of targeted A• T to G• C through an intermediated that reads as guanine (G) by polymerases during DNA replication. 2 Koblan et al. used an optimized ABE7. 10 variant (ABEmax-VRQR) 3 with sgRNA targeting c. 1824 C> T in LMNA (Fig. 1 b). 1 Through the lentiviral delivery, Koblan et al. tested the ABEmax-VRQR in two primary cells derived from HGPS patients, resulting in the correction of~ 90% of the mutated genes. 1 This led to normal splicing of laminA, reduced a notable expression and accumulation of progerin, and substantially corrected the abnormal nucleus (Fig. 1 b). 1 Thus, the ABE approach developed in this study could repair the mutated gene in HGPS cells.Next, to investigate the use of ABE treatment to correct the HGPS mutation in vivo, Koblan et al. used the clinical adenoassociated virus vector for co-packaged ABEmax-VRQR and sgRNA (ABE-AAV) to deliver the repair complex machinery into the HGPS mouse model (Fig. 1 c) 1—C57BL/6 mice homozygous includes the pathogenic human LMNA c. 1824 C> T allele that develops many hallmark symptoms seen in human HGPS patients, such as loss of vascular smooth muscle cells (VSMCs), cardiovascular complication, loss of subcutaneous fat and early death. 4 Following the retro-orbital injection of ABE-AAV in mice at postnatal day 3 (P3) and 14 (P14), Koblan et al. achieved a 10–60% correction of the human …
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一项对患有 A 型血友病的狗进行 AAV 基因治疗的长期研究确定了转导肝细胞的克隆扩增。
影响因子：
46.9
作者：
Nguyen GN;Everett JK;Kafle S;Roche AM;Raymond HE;Leiby J;Wood C;Assenmacher CA;Merricks EP;Long CT;Kazazian HH;Nichols TC;Bushman FD;Sabatino DE
通讯作者：
Sabatino DE
DOI：
10.1073/pnas.0600012103
发表时间：
2006-02-28
影响因子：
11.1
作者：
Varga, R;Eriksson, M;Collins, FS
通讯作者：
Collins, FS
影响因子：
64.8
作者：
Koblan LW;Erdos MR;Wilson C;Cabral WA;Levy JM;Xiong ZM;Tavarez UL;Davison LM;Gete YG;Mao X;Newby GA;Doherty SP;Narisu N;Sheng Q;Krilow C;Lin CY;Gordon LB;Cao K;Collins FS;Brown JD;Liu DR
通讯作者：
Liu DR
影响因子：
46.9
作者：
Huang, Tony P.;Zhao, Kevin T.;Liu, David R.
通讯作者：
Liu, David R.