pLenti-EIF3A-sgRNA (EIF3A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EIF3A基因敲除的质粒(L02600 pLenti-EIF3A-sgRNA)、慢病毒(L02601 EIF3A Knockout Lentivirus)、HEK293T细胞(L02602 EIF3A Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L02603 EIF3A Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L02604 EIF3A 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。
EIF3A基因的基本信息如下:
Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
Human | EIF3A | 8661 | BC114429 | NM_003750 |
About the gene | |
Official Symbol | EIF3A |
Previous Symbol | EIF3|EIF3S10 |
Official Full Name | eukaryotic translation initiation factor 3 subunit A |
Synonyms | eIF3-theta; eIF3-p170; KIAA0139; eIF3a; TIF32 |
Location | 10q26.11 |
Gene Type | protein-coding gene |
Uniprot ID | Q14152 |
Pathway/Library | m6A Modification Related Genes Library |
Gene Summary | RNA-binding component of the eukaryotic translation initiation factor 3 (eIF-3) complex, which is required for several steps in the initiation of protein synthesis (PubMed:17581632, PubMed:25849773). The eIF-3 complex associates with the 40S ribosome and facilitates the recruitment of eIF-1, eIF-1A, eIF-2:GTP:methionyl-tRNAi and eIF-5 to form the 43S pre-initiation complex (43S PIC). The eIF-3 complex stimulates mRNA recruitment to the 43S PIC and scanning of the mRNA for AUG recognition. The eIF-3 complex is also required for disassembly and recycling of post-termination ribosomal complexes and subsequently prevents premature joining of the 40S and 60S ribosomal subunits prior to initiation (PubMed:17581632, PubMed:11169732). The eIF-3 complex specifically targets and initiates translation of a subset of mRNAs involved in cell proliferation, including cell cycling, differentiation and apoptosis, and uses different modes of RNA stem-loop binding to exert either translational activation or repression (PubMed:25849773, PubMed:27462815). EIF3A_HUMAN,Q14152 (Microbial infection) Essential for the initiation of translation on type-1 viral ribosomal entry sites (IRESs), like for HCV, PV, EV71 or BEV translation (PubMed:23766293, PubMed:24357634). EIF3A_HUMAN,Q14152 (Microbial infection) In case of FCV infection, plays a role in the ribosomal termination-reinitiation event leading to the translation of VP2 (PubMed:18056426). EIF3A_HUMAN,Q14152 |
产品编号 | 产品名称 | 包装 |
L02600 | pLenti-EIF3A-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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