pLenti-TRIM67-sgRNA (TRIM67基因敲除质粒)是一种在动物细胞中可以同时表达Cas9、目的基因的sgRNA和puromycin抗性基因的质粒。用于在动物细胞中直接基于CRISPR/Cas9技术敲除目的基因,或者通过包装慢病毒后基于CRISPR/Cas9技术敲除目的基因。本质粒中sgRNA的有效性已经通过T7EI法的验证。
本质粒在细菌中为Amp抗性,全长约13,000bp。本质粒的关键图谱信息请参考图1。本质粒可直接转染细胞用于目的基因的CRISPR/Cas9敲除,以及通过puromycin筛选稳定细胞株;也可以与pMDLg、Rev及VSV-g共转HEK293T细胞进行重组慢病毒(lentivirus)的包装,然后再用于感染细胞或组织并进行目的基因的CRISPR/Cas9敲除。
图1. 表达sgRNA、Cas9和puromycin抗性的pLenti-sgRNA质粒关键图谱信息。
本质粒中的sgRNA基于碧云天研发的CRISPR/Cas9 sgRNA快速筛选和验证体系获得,sgRNA的有效性已经通过T7EI法验证。
本质粒用于实验时,建议同时选购无任何靶向的对照质粒pLenti-Control-sgRNA (L00011)或靶向GFP的对照质粒pLenti-GFP-sgRNA (L00013)。
碧云天同时提供基于CRISPR/Cas9技术的TRIM67基因敲除的质粒(L10975 pLenti-TRIM67-sgRNA)、慢病毒(L10976 TRIM67 Knockout Lentivirus)、HEK293T细胞(L10977 TRIM67 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L10978 TRIM67 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L10979 TRIM67 Knockout HEK293T Trizol Lysate)等产品,具体请在碧云天网站查询或在本产品网页点击相应产品。
TRIM67基因的基本信息如下:
Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
Human | TRIM67 | 440730 | - | NM_001004342 |
About the gene | |
Official Symbol | TRIM67 |
Previous Symbol | - |
Official Full Name | tripartite motif containing 67 |
Synonyms | TNL |
Location | 1q42.2 |
Gene Type | protein_coding |
Uniprot ID | Q6ZTA4.3 |
Pathway/Library | Ubiquitin Ligases Genes Library |
Gene Summary | Tripartite motif (TRIM) proteins have been increasingly appreciated as important antiviral factors that suppress the replication of a wide range of RNA and DNA viruses. TRIM proteins inhibit viral replication by either directly targeting viral components or modulating innate immune responses that result in antiviral gene expression. For example, TRIM19 (also known as promyelocytic leukemia protein (PML)) restricts multiple RNA viruses and DNA viruses, including herpesviruses, by organizing PML nuclear bodies. Several TRIM proteins that suppress KSHV reactivation. TRIM67, was commonly silenced in colorectal cancer and its downregulation was associated with poor survival. Trim67 knockout in ApcMin/+ mice increased the incidence, multiplicity, and burden of colorectal tumors. Similarly, colon-specific knockout of Trim67 significantly accelerated azoxymethane-induced colorectal cancer in mice. RNA sequencing revealed that the antitumor effect of TRIM67 was mediated by activation of the p53 signaling pathway. TRIM67 interacted directly with the C-terminus of p53, inhibiting p53 degradation by its ubiquitin ligase MDM2. TRIM67 was also a transcriptional target of p53; upon cellular stress, p53 bound to the TRIM67 promoter and induced significant upregulation of TRIM67, thereby forming a TRIM67/p53 self-amplifying loop that boosts p53-induced cell growth inhibition and apoptosis. Consequently, loss of this p53-positive regulatory program profoundly compromised p53-mediated responses to chemotherapy-induced DNA damage. Dampened p53 response was also observed in tumors of Trim67 knockout mice and Trim67 knockout embryonic fibroblasts. TRIM67 reactivation restored p53 activation and sensitized colorectal cancer cells to chemotherapy in vitro and in vivo. TRIM67 thus functions as a pivotal tumor suppressor in colorectal cancer and is a potential target for improving chemotherapy responsiveness. |
产品编号 | 产品名称 | 包装 |
L10975 | pLenti-TRIM67-sgRNA | 5µg |
— | 说明书 | 1份 |
-20℃保存,至少两年有效。
注意事项:碧云天拥有sgRNA序列的知识产权,如果需要sgRNA序列,请在订购后发送邮件向info@beyotime.com索取。sgRNA与质粒及其序列信息,未经碧云天书面许可不得用于任何商业用途,也不得移交给订货人所在实验室外的任何个人或单位。使用者在发表研究论文或结果时,应注明来源。
慢病毒包装使用的包装质粒,可以订购碧云天的Lentivirus Packaging Vectors Set A (L00002),包括pMDLg、Rev和VSV-g。
对于非目录产品的CRISPR基因敲除用的sgRNA表达质粒的定制,可联系碧云天技术服务service@beyotime.com。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
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