2023年12月12日 · By engineering TREX2 to degrade 3′ overhangs generated by paired Cas9ns, the corresponding author and colleagues develop a method to stimulate paired Cas9n-induced genome disruption up to 400-fold, thus expanding the utility of paired Cas9n in genome editing.

2015年7月13日 · As in the TREX2 experiments, 500,000 293T cells were seeded, and approximately 24 h later, 2 μg of sgRNA and 2 μg of Cas9 Sp (or 2 μg of Cas9 St1) were cotransfected using Lipofectamine 3000 at...

A modified calcium phosphate (CaPO 4) transfection protocol 31 was used to introduce a pRRL-based lentiviral expression vector encoding mCreI, TREX2, and mCherry proteins into human 293T cells 32 (Supplementary Fig. S1). Cells (2–4 × 10 5 /well) were plated on a six-well plate 24 h prior and grown to ∼70% confluence.

TERM2, full named as Triggering receptor expressed on myeloid cells 2, is a membrane protein which expressed primarily in immune cells across many different tissues, and form a receptor signaling complex with TYROBP, also named as DAP12, then mediates signaling and cell activation after binding serval ligands, it plays an important role in diffe...

expression and TREX2 fusion on repair of 30-overhanging ends generated by paired SpCas9n on non-overlapping and overlapping targets. We found that both Trex2 co-expression and TREX2 fusion stimulate the efficiency of paired SpCas9n in genome disruption with 30-overhanging ends. In particular, the SpCas9n-TREX2 fusion

Plasmids encoding TREX2 (#40210), Artemis (#40211), and empty backbone (#39991) were obtained from Addgene. Given the high probability of a single cell achieving multiple NHEJ events due to the presence of multiple target sites, DNA from cells are harvested 72hrs post-transfection to minimize loss of cells with large amounts of nuclease activity.

2016年2月2日 · We demonstrate a simple droplet digital PCR assay that quickly quantitates a range of indel mutations with detection as low as 0.02% mutant in a wild type background and precision (≤6%CV) and...

通过研究者两个质粒的backbone以及文章描述，我们大概可以明白：（1）293T细胞具有SV40 T区域，使得带有SV40 ori质粒能够快速扩增；（2）在293T细胞中转入带有逆转录病毒元件+半乳糖苷酶(B-galactosidase)-SV40 promoter的质粒，结合SV40大T抗原，从此逆转录病毒产量得以 ...

T-REx™ 细胞系可稳定表达四环素抑制子蛋白（表 1）。 使用 T-REx™ 系统可以节省大量的时间和精力。 通过使用阳性对照载体 pcDNA™4⁄TO⁄lacZ 了解更多信息. Have Questions? T-REx™ 细胞系可稳定表达四环素抑制子蛋白（表 1）。 使用 T-REx™ 系统可以节省大量的时间和精力。 通过使用阳性对照载体 pcDNA™4⁄TO⁄ lac Z 进行瞬时转染，对 T-REx™ 细胞系进行功能检测。 在用四环素诱导时，T-REx™ 细胞系在抑制状态下表现出极低的基础表达水平，表达水平较高（图 …

HEK-293T细胞是293T（293tsA1609neo）细胞系（ATCC CRL-11268）的衍生物。细胞持续表达SV40抗原，该细胞常用于转染实验，转染效率较高。（转染一般以50%密度铺板，待细胞刚刚贴到皿壁上后（一般8-10小时），即可进行转染操作)。