细胞转染注意事项,你都get了吗？(下)
细胞转染实验从开始设计到最终的效果检测，可以由6个过程组成：核酸准备、试剂选择、细胞铺板、转染操作、细胞培养、效果检测。每一步都会影响最终的结果，均存在不同的注意事项。上个月小翌分享了前3个过程的注意事项：
本期分享后面3个过程的注意事项：
转染操作
1.条件摸索与优化：
2.无血清配置复合物：
3.孵育时间：
4.抗生素影响：
1.优良的培养试剂：
2.是否换液：
1.检测方法的选择：
2.检测时间的选择：
3.稳转筛选：
[1] Liu R, Yang J, et al. Optogenetic control of RNA function and metabolism using engineered light-switchable RNA-binding proteins. Nat Biotechnol. 2022 Jan 3. (IF:55)华东理工大学生物工程学院杨弋团队
[2] Zhou J, Chen P, et al. Cas12a variants designed for lower genome-wide off-target effect through stringent PAM recognition. Mol Ther. 2022 Jan 5.(IF:11.454)武汉大学生命科学学院殷雷团队
[3] Chen S, Cao X, et al. circVAMP3 Drives CAPRIN1 Phase Separation and Inhibits Hepatocellular Carcinoma by Suppressing c-Myc Translation. Adv Sci (Weinh). 2022 Jan 24.(IF:16.808)中国科学院北京生命科学研究院赵方庆团队
[4] Gu C, Wang Y, et al. AHSA1 is a promising therapeutic target for cellular proliferation and proteasome inhibitor resistance in multiple myeloma. J Exp Clin Cancer Res. 2022 Jan 6.(11.161)南京中医药大学杨烨顾春艳团队
[5] Zhang Y, Yu X, et al. Splicing factor arginine/serine-rich 8 promotes multiple myeloma malignancy and bone lesion through alternative splicing of CACYBP and exosome-based cellular communication. Clin Transl Med. 2022 Feb.(11.492)南京中医药大学杨烨顾春艳团队
[6] Qin J, Cai Y, et al. Molecular mechanism of agonism and inverse agonism in ghrelin receptor. Nat Commun. 2022 Jan 13.(14.9)四川大学生物治疗国家重点实验室邵振华团队
[7] Tang X, Deng Z, et al. A novel protein encoded by circHNRNPU promotes multiple myeloma progression by regulating the bone marrow microenvironment and alternative splicing. J Exp Clin Cancer Res. 2022 Mar 8.(11.161)南京中医药大学杨烨顾春艳团队
[8] Xie F, Su P, et al. Engineering Extracellular Vesicles Enriched with Palmitoylated ACE2 as COVID-19 Therapy. Adv Mater. 2021 Oct 19. (IF:30.849)苏州大学生物医学研究院周芳芳团队和浙江大学生命科学研究院张龙团队
[9] Liang Y, Lu Q, et al. Reactivation of tumour suppressor in breast cancer by enhancer switching through NamiRNA network. Nucleic Acids Res. 2021 Sep 7.(IF:16.9)复旦大学生物医学研究院于文强团队
[10] Fan Y, Wang J, et al. CircNR3C2 promotes HRD1-mediated tumor-suppressive effect via sponging miR-513a-3p in triple-negative breast cancer. Mol Cancer. 2021 Feb 2.(IF:27.403)南京医科大学附属逸夫医院苏东明团队
[11] Dai L, Dai Y, et al. Structural insight into BRCA1-BARD1 complex recruitment to damaged chromatin. Mol Cell. 2021 Jul 1.(IF:17.97)浙江大学生命科学研究院黄俊团队和中科院生物物理所周政团队
[12] Zhang K, Wang A, et al. UBQLN2-HSP70 axis reduces poly-Gly-Ala aggregates and alleviates behavioral defects in the C9ORF72 animal model. Neuron. 2021 Jun 16.(IF:17.17)中国科学院生物与化学交叉研究中心王文元团队
[13] Li T, Chen X, et al. A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice. Nat Commun. 2021 Jan 27.(IF:14.92)华东理工大学生物工程学院杨弋团队
[14] Pan Y, He X, et al. Neuronal activity recruits the CRTC1/CREB axis to drive transcription-dependent autophagy for maintaining late-phase LTD. Cell Rep. 2021 Jul 20.(IF:9.420)浙江大学脑科学与脑医学学院马欢团队
[15] Liu H, Xing R, et al. G-protein-coupled receptor GPR17 inhibits glioma development by increasing polycomb repressive complex 1-mediated ROS production. Cell Death Dis. 2021 Jun 12.(IF:8.463)厦门大学生命科学学院陈颖团队
[16] Yan F, Huang C, et al. Threonine ADP-Ribosylation of Ubiquitin by a Bacterial Effector Family Blocks Host Ubiquitination. Mol Cell. 2020 May 21.(IF:17.97)浙江大学生命科学研究院朱永群团队
[17] Luo Q, Wu X, et al. TRIM32/USP11 Balances ARID1A Stability and the Oncogenic/Tumor-Suppressive Status of Squamous Cell Carcinoma. Cell Rep. 2020 Jan 7.(IF:9.42)中国医学科学院分子肿瘤学国家重点实验室刘芝华团队
[18] Sun X, Peng X, et al. ADNP promotes neural differentiation by modulating Wnt/β-catenin signaling. Nat Commun. 2020 Jun 12.(IF:14.911)中国科学院水生生物研究所孙玉华团队
[19] Yang X, Wang H, et al. Rewiring ERBB3 and ERK signaling confers resistance to FGFR1 inhibition in gastrointestinal cancer harbored an ERBB3-E928G mutation. Protein Cell. 2020 Dec.(IF:14.872)浙江大学医学院/转化医学研究院闵军霞团队
[20] Zou Y, Wang A, et al. Analysis of redox landscapes and dynamics in living cells and in vivo using genetically encoded fluorescent sensors. Nat Protoc. 2018 Oct.(IF:13.490)华东理工大学生物工程学院杨弋、赵玉政团队
[21] Zhang K, Zhao X, et al. Enhanced Therapeutic Effects of Mesenchymal Stem Cell-Derived Exosomes with an Injectable Hydrogel for Hindlimb Ischemia Treatment. ACS Appl Mater Interfaces. 2018 Sep 12.(IF:8.09)南开大学医学院李宗金团队
[22] Hao H, Hu S, et al. Loss of Endothelial CXCR7 Impairs Vascular Homeostasis and Cardiac Remodeling After Myocardial Infarction: Implications for Cardiovascular Drug Discovery. Circulation. 2017 Mar 28.(IF:29.69)中国医学科学院/北京协和医学院阜外医院王淼团队
|
应用场景 |
名称 |
货号 |
规格 |
|
细胞类型：贴壁/悬浮 核酸类型：DNA、siRNA |
40802ES02 |
0.5 mL |
|
|
40802ES03 |
1.0 mL |
||
|
40802ES08 |
5×1 mL |
||
|
细胞类型：贴壁/悬浮 |
40803ES70 |
200 T |
|
|
用途：病毒感染、DNA转染 |
40804ES76 |
500 μL |
|
|
40804ES86 |
5×500 μL |
||
|
细胞类型：悬浮 |
40805ES02 |
0.5 mL |
|
|
40805ES03 |
1.0 mL |
||
|
40805ES08 |
5×1 mL |
||
|
细胞类型：贴壁/悬浮 |
40806ES01 |
0.1 mL |
|
|
40806ES02 |
0.5 mL |
||
|
40806ES03 |
1.0 mL |
||
|
细胞类型：贴壁/悬浮 |
40808ES02 |
0.5 mL |
|
|
40808ES03 |
1 mL |
||
|
40808ES08 |
5×1 mL |
||
|
细胞类型：贴壁/悬浮 |
40809ES01 |
0.1 mL |
|
|
40809ES03 |
1 mL |
||
|
细胞类型：贴壁/悬浮 |
40815ES03 |
1 g |
|
|
40815ES08 |
5×1 g |
||
|
细胞类型：贴壁/悬浮 |
40816ES02 |
100 mg |
|
|
40816ES03 |
1 g |
||
|
细胞类型：293 |
40820ES04 |
1.5 mL |
|
|
40820ES10 |
10 mL |
||
|
40820ES60 |
100 mL |
||
|
细胞类型：293 核酸类型：DNA 用途：AAV/LV载体大规模生产 |
40821ES10 |
10 mL |
|
|
40821ES60 |
100 mL |
||
|
40821ES80 |
1 L |
