RC3H2 Knockout HEK293T Cells (RC3H2基因敲除HEK293T细胞)是通过同时表达Cas9、目的基因sgRNA和puromycin抗性基因,并实现了目的基因CRISPR敲除的HEK293T细胞。本细胞中目的基因的敲除已经通过T7EI法的验证。本细胞株是多克隆细胞,可用于该目的基因的生物学功能研究,也可以用于该基因相应抗体的验证。
本HEK293T Cells为可同时表达Cas9、puromycin抗性基因和目的基因sgRNA的慢病毒感染HEK293T细胞并经过puromycin筛选后获得的多克隆HEK293T细胞。制备本细胞的相应慢病毒的基因序列的关键图谱信息请参考图1。
图1. 可同时表达sgRNA、Cas9和puromycin抗性的慢病毒其基因序列的关键图谱信息。
本细胞中目的基因的敲除已经通过T7EI法的验证。
由于本细胞是通过CRISPR/Cas9技术获得的多克隆细胞,基于CRISPR/Cas9技术的特点,理论上平均有2/3的细胞发生移码突变而导致了目的基因的敲除,平均有1/3的细胞并未发生移码突变。很多情况下有约2/3的细胞发生目的基因的敲除,已经足以进行很多的目的基因的生物学功能的研究了。如果希望获得100%基因敲除的细胞,可以自行使用本产品筛选单克隆细胞,或者委托碧云天进行单克隆细胞株的筛选服务。
本细胞用于实验时,建议同时选购无任何靶向的对照细胞Control Knockout HEK293T Cells (L00020)或靶向GFP的对照细胞GFP Knockout HEK293T Cells (L00022)。
碧云天同时提供基于CRISPR/Cas9技术的RC3H2基因敲除的质粒(L10115 pLenti-RC3H2-sgRNA)、慢病毒(L10116 RC3H2 Knockout Lentivirus)、HEK293T细胞(L10117 RC3H2 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L10118 RC3H2 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L10119 RC3H2 Knockout HEK293T Trizol Lysate)等产品等产品,具体请在碧云天网站查询或在本产品网页点击相应产品。
RC3H2基因的基本信息如下:
Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
Human | RC3H2 | 54542 | BC044642 | NM_018835 |
About the gene | |
Official Symbol | RC3H2 |
Previous Symbol | MNAB |
Official Full Name | ring finger and CCCH-type domains 2 |
Synonyms | FLJ20301; FLJ20713; RNF164 |
Location | 9q33.2 |
Gene Type | protein-coding gene |
Uniprot ID | Q9HBD1 |
Pathway/Library | Ubiquitin Ligases Genes Library |
Gene Summary | Post-transcriptional repressor of mRNAs containing a conserved stem loop motif, called constitutive decay element (CDE), which is often located in the 3'-UTR, as in HMGXB3, ICOS, IER3, NFKBID, NFKBIZ, PPP1R10, TNF and in many more mRNAs. Binds to CDE and promotes mRNA deadenylation and degradation. This process does not involve miRNAs. In follicular helper T (Tfh) cells, represses of ICOS and TNFRSF4 expression, thus preventing spontaneous Tfh cell differentiation, germinal center B-cell differentiation in the absence of immunization and autoimmunity. In resting or LPS-stimulated macrophages, controls inflammation by suppressing TNF expression. Also recognizes CDE in its own mRNA and in that of paralogous RC3H1, possibly leading to feedback loop regulation (By similarity). miRNA-binding protein that regulates microRNA homeostasis. Enhances DICER-mediated processing of pre-MIR146a but reduces mature MIR146a levels through an increase of 3' end uridylation. Both inhibits ICOS mRNA expression and they may act together to exert the suppression (PubMed:25697406). Acts as a ubiquitin E3 ligase. Pairs with E2 enzymes UBE2B, UBE2D2, UBE2E2, UBE2E3, UBE2G2, UBE2K and UBE2Q2 and produces polyubiquitin chains (PubMed:26489670). Shows the strongest activity when paired with UBE2N:UBE2V1 or UBE2N:UBE2V2 E2 complexes and generate both short and long polyubiquitin chains (PubMed:26489670). Involved in the ubiquitination of MAP3K5 (PubMed:24448648, PubMed:26489670) (By similarity). Able to interact with double-stranded RNA (dsRNA) (PubMed:26489670). RC3H2_HUMAN,Q9HBD1 |
产品编号 | 产品名称 | 包装 |
L10117 | RC3H2 Knockout HEK293T Cells | 1支/瓶 |
— | 说明书 | 1份 |
对于细胞培养瓶或离心管运输的活细胞,室温3-5天有效;对于干冰运输的冻存细胞,液氮保存,长期有效。
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序列信息与本细胞,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。
对于本细胞的单克隆细胞株筛选或非HEK293T细胞的CRISPR基因敲除细胞的定制,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。
为了您的安全和健康,请穿实验服并戴一次性手套操作。
产品编号 | 产品名称 | 包装 |
L00020 | Control Knockout HEK293T Cells | 1支/瓶 |
L00022 | GFP Knockout HEK293T Cells | 1支/瓶 |
C0222 | 青霉素-链霉素溶液(100X) | 100ml |
D0508S/M | 基因组编辑突变检测试剂盒 | 25/100次 |
D7080S/M/L | T7 Endonuclease I (CRISPR等基因突变鉴定用) | 250/1250/5000U |
ST551-10mg | Puromycin Dihydrochloride (嘌呤霉素) | 10mg/ml×1ml |
ST551-50mg | Puromycin Dihydrochloride (嘌呤霉素) | 10mg/ml×5ml |
ST551-250mg | Puromycin Dihydrochloride (嘌呤霉素) | 250mg |