Targeted delivery of CRISPR-Cas9 and transgenes enables complex immune cell engineering.
Targeted delivery of CRISPR-Cas9 and transgenes enables complex immune cell engineering.
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CRISPR-Cas9和转基因的靶向递送实现了复杂的免疫细胞工程。
DOI：
10.1016/j.celrep.2021.109207
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发表时间：
2021-06-01
期刊：
影响因子：
8.8
通讯作者：
Doudna JA
中科院分区：
文献类型：
--
作者：
Hamilton JR;Tsuchida CA;Nguyen DN;Shy BR;McGarrigle ER;Sandoval Espinoza CR;Carr D;Blaeschke F;Marson A;Doudna JA
随着基因组工程推进基于细胞的疗法，将CRISPR-Cas9核糖核蛋白（RNP）和治疗性转基因引入特定细胞的通用方法将是变革性的。通过病毒转导制造的表达嵌合抗原受体（CAR）的自体T细胞被批准用于治疗多种血癌，但可能需要额外的遗传修饰来改变细胞程序以治疗实体瘤和同种异体细胞疗法。我们已经开发了一种一步策略，使用工程化的慢病毒颗粒将Cas9 RNP和CAR转基因引入原代人T细胞而无需电穿孔。此外，编程颗粒向性允许我们靶向混合细胞群中的特定细胞类型。作为概念验证，我们表明HIV-1包膜靶向颗粒编辑CD 4+细胞，同时保留共培养的CD 8+细胞。这种离体免疫细胞工程的适应性方法提供了适用于体内靶向体细胞的遗传修饰的策略。汉密尔顿等人证明，工程化的病毒样颗粒能够同时进行CRISPR-Cas9基因敲除和T细胞的抗肿瘤重编程。通过向外制造类似于HIV-1的病毒样颗粒，作者还证明了在混合细胞群中特异性靶向CD 4 + T细胞进行基因组编辑。
As genome engineering advances cell-based therapies, a versatile approach to introducing both CRISPR-Cas9 ribonucleoproteins (RNPs) and therapeutic transgenes into specific cells would be transformative. Autologous T cells expressing a chimeric antigen receptor (CAR) manufactured by viral transduction are approved to treat multiple blood cancers, but additional genetic modifications to alter cell programs will likely be required to treat solid tumors and for allogeneic cellular therapies. We have developed a one-step strategy using engineered lentiviral particles to introduce Cas9 RNPs and a CAR transgene into primary human T cells without electroporation. Furthermore, programming particle tropism allows us to target a specific cell type within a mixed cell population. As a proof-of-concept, we show that HIV-1 envelope targeted particles to edit CD4+ cells while sparing co-cultured CD8+ cells. This adaptable approach to immune cell engineering ex vivo provides a strategy applicable to the genetic modification of targeted somatic cells in vivo. Hamilton et al. demonstrate that engineered virus-like particles enable simultaneous CRISPR-Cas9 genetic knockout and the anti-tumor reprogramming of T cells. By outwardly making virus-like particles resemble HIV-1, the authors additionally demonstrate specific targeting of CD4+ T cells for genome editing within a mixed cell population.
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影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
7
作者：
Ramakrishna S;Kwaku Dad AB;Beloor J;Gopalappa R;Lee SK;Kim H
通讯作者：
Kim H
DOI：
10.1038/nrc.2016.97
发表时间：
2016-08-23
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
Fesnak AD;June CH;Levine BL
通讯作者：
Levine BL
影响因子：
14.8
作者：
Hill, Zachary B.;Martinko, Alexander J.;Wells, James A.
通讯作者：
Wells, James A.
影响因子：
7.3
作者：
Muller YD;Nguyen DP;Ferreira LMR;Ho P;Raffin C;Valencia RVB;Congrave-Wilson Z;Roth TL;Eyquem J;Van Gool F;Marson A;Perez L;Wells JA;Bluestone JA;Tang Q
通讯作者：
Tang Q