RC3H2 Knockout HEK293T Trizol Lysate (RC3H2基因敲除HEK293T细胞Trizol裂解液)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敲除的多克隆HEK293T细胞的Trizol裂解液。该细胞中目的基因的敲除已经通过T7EI法的验证。本产品可用于该目的基因敲除后其信号通路相关RNA表达的研究。
本Trizol裂解液源于可同时表达Cas9、目的基因sgRNA和puromycin抗性基因的慢病毒感染HEK293T细胞并经过puromycin筛选后获得的多克隆HEK293T细胞。制备该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。
该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Trizol裂解液用于实验时，建议同时选购无任何靶向的对照细胞Trizol裂解液Control Knockout HEK293T Trizol Lysate (L00031)或靶向GFP的对照Trizol裂解液GFP Knockout HEK293T Trizol Lysate (L00033)。
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，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 |
| L10119 | RC3H2 Knockout HEK293T Trizol Lysate | 500µl |
| — | 说明书 | 1份 |
-20℃保存，至少6个月有效；-80℃保存，至少一年有效。
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序列信息与本产品，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。
对于非目录产品的CRISPR基因敲除细胞Trizol裂解液的定制，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 |
| L00031 | Control Knockout HEK293T Trizol Lysate | 500µl |
| L00033 | GFP Knockout HEK293T Trizol Lysate | 500µl |
| D7166 | BeyoRT™ cDNA第一链合成试剂盒(RNase H-) | 10次 |
| D7168 | BeyoRT™ II cDNA第一链合成试剂盒(RNase H-) | 20/100/500次 |
| D7170 | BeyoRT™ II cDNA合成试剂盒(with gDNA Eraser) | 20/100/500次 |
| D7178 | BeyoRT™ III cDNA第一链合成试剂盒 | 20/100/500次 |
| D7180 | BeyoRT™ III cDNA合成试剂盒 (with gDNA EZeraser) | 20/100/500次 |
| D7182 | BeyoRT™ III cDNA第一链合成预混液(5X) | 20/100/500次 |
| D7185 | BeyoRT™ III cDNA合成预混液(5X)(with gDNA EZeraser) | 20/100/500次 |
| D7260 | BeyoFast™ SYBR Green qPCR Mix (2X) | 1/5/25ml |
| D7262 | BeyoFast™ SYBR Green qPCR Mix (2X, Low ROX) | 1/5/25ml |
| D7265 | BeyoFast™ SYBR Green qPCR Mix (2X, High ROX) | 1/5/25ml |
| D7268 | BeyoFast™ SYBR Green One-Step qRT-PCR Kit | 100/500次 |
| D7271 | BeyoFast™ Probe qPCR Mix (2X) | 1/5/25ml |
| D7272 | BeyoFast™ Probe qPCR Mix (2X, Low ROX) | 1/5/25ml |
| D7273 | BeyoFast™ Probe qPCR Mix (2X, High ROX) | 1/5/25ml |
| D7277 | BeyoFast™ Probe One-Step qRT-PCR Kit | 100/500次 |
| R0011 | Beyozol (总RNA抽提试剂) | 100ml |
| R0016 | Trizol (总RNA抽提试剂) | 100ml |
