In vivo genome editing at the albumin locus to treat methylmalonic acidemia.
In vivo genome editing at the albumin locus to treat methylmalonic acidemia.
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DOI：
10.1016/j.omtm.2021.11.004
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发表时间：
2021-12-10
期刊：
影响因子：
--
通讯作者：
Venditti CP
中科院分区：
文献类型：
--
作者：
Schneller JL;Lee CM;Venturoni LE;Chandler RJ;Li A;Myung S;Cradick TJ;Hurley AE;Lagor WR;Bao G;Venditti CP
甲基丙二酸血症（MMA）是一种代谢紊乱，最常见的甲基丙二酰辅酶A β（MMUT）基因突变引起的。虽然腺相关病毒（AAV）基因治疗已有效地纠正MMA小鼠模型中的疾病表型，但临床翻译可能会因附加型转基因表达的丧失而受损，并因需要在生命早期治疗患者而扩大。为了实现永久校正，我们开发了一种双重AAV策略来表达来自Alb的密码子优化的MMUT转基因，并在新开发的MMA敲入小鼠模型中测试了各种CRISPR-Cas9基因组编辑载体。对于白蛋白内含子1中的一个靶位点，我们设计了在2A肽或内部核糖体进入位点序列后面表达MMUT的救援盒。第二个指导RNA靶向起始密码子，供体盒包含5′同源臂中的近端白蛋白启动子。尽管所有编辑方法都是治疗性的，但靶向白蛋白的起始密码子允许使用也作为附加体和同源重组后的供体盒，即使没有Cas9的表达，也作为整合体。使用这些策略靶向白蛋白位点将对需要早期治疗和永久性长期纠正的其他代谢疾病有效。甲基丙二酸血症，MMUT亚型，是一种严重的代谢紊乱，通过选择性肝移植治疗。我们开发了几种治疗性基因组编辑方法，使用和不使用核酸酶，使用AAV载体将MMUT基因插入白蛋白基因座的5′端。我们的研究结果可广泛推广到其他代谢性疾病。
Methylmalonic acidemia (MMA) is a metabolic disorder most commonly caused by mutations in the methylmalonyl-CoA mutase (MMUT) gene. Although adeno-associated viral (AAV) gene therapy has been effective at correcting the disease phenotype in MMA mouse models, clinical translation may be impaired by loss of episomal transgene expression and magnified by the need to treat patients early in life. To achieve permanent correction, we developed a dual AAV strategy to express a codon-optimized MMUT transgene from Alb and tested various CRISPR-Cas9 genome-editing vectors in newly developed knockin mouse models of MMA. For one target site in intron 1 of Alb, we designed rescue cassettes expressing MMUT behind a 2A-peptide or an internal ribosomal entry site sequence. A second guide RNA targeted the initiator codon, and the donor cassette encompassed the proximal albumin promoter in the 5′ homology arm. Although all editing approaches were therapeutic, targeting the start codon of albumin allowed the use of a donor cassette that also functioned as an episome and after homologous recombination, even without the expression of Cas9, as an integrant. Targeting the albumin locus using these strategies would be effective for other metabolic disorders where early treatment and permanent long-term correction are needed. Methylmalonic acidemia, MMUT subtype, is a severe metabolic disorder treated by elective liver transplantation. We developed several therapeutic genome-editing approaches, with and without nucleases, to insert the MMUT gene into the 5′ end of the albumin locus using AAV vectors. Our results should be broadly extendable other metabolic disorders.
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影响因子：
15.9
作者：
Chandler, Randy J.;LaFave, Matthew C.;Venditti, Charles P.
通讯作者：
Venditti, Charles P.
DOI：
10.1016/j.ymthe.2017.09.020
发表时间：
2017-11-01
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
Borel F;Tang Q;Gernoux G;Greer C;Wang Z;Barzel A;Kay MA;Shultz LD;Greiner DL;Flotte TR;Brehm MA;Mueller C
通讯作者：
Mueller C
影响因子：
64.8
作者：
Barzel, A.;Paulk, N. K.;Shi, Y.;Huang, Y.;Chu, K.;Zhang, F.;Valdmanis, P. N.;Spector, L. P.;Porteus, M. H.;Gaensler, K. M.;Kay, M. A.
通讯作者：
Kay, M. A.
影响因子：
3.8
作者：
Chandler, Randy J.;Venditti, Charles P.
通讯作者：
Venditti, Charles P.
DOI：
10.1073/pnas.1302764110
发表时间：
2013-08-13
影响因子：
11.1
作者：
Manoli, Irini;Sysol, Justin R.;Venditti, Charles P.
通讯作者：
Venditti, Charles P.