CCAR2 Knockout HEK293T Trizol Lysate (CCAR2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CCAR2基因敲除的质粒(L24975 pLenti-CCAR2-sgRNA)、慢病毒(L24976 CCAR2 Knockout Lentivirus)、HEK293T细胞(L24977 CCAR2 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L24978 CCAR2 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L24979 CCAR2 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。
CCAR2基因的基本信息如下：
| Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
| Human | CCAR2 | 57805 | BC018269 | NM_021174 |
| About the gene | |
| Official Symbol | CCAR2 |
| Previous Symbol | KIAA1967 |
| Official Full Name | cell cycle and apoptosis regulator 2 |
| Synonyms | DBC-1; DBC1; NET35 |
| Location | 8p21.3 |
| Gene Type | protein-coding gene |
| Uniprot ID | Q8N163 |
| Pathway/Library | others |
| Gene Summary | Core component of the DBIRD complex, a multiprotein complex that acts at the interface between core mRNP particles and RNA polymerase II (RNAPII) and integrates transcript elongation with the regulation of alternative splicing: the DBIRD complex affects local transcript elongation rates and alternative splicing of a large set of exons embedded in (A + T)-rich DNA regions (PubMed:22446626). Inhibits SIRT1 deacetylase activity leading to increasing levels of p53/TP53 acetylation and p53-mediated apoptosis (PubMed:18235501, PubMed:18235502, PubMed:23352644). Inhibits SUV39H1 methyltransferase activity (PubMed:19218236). Mediates ligand-dependent transcriptional activation by nuclear hormone receptors (PubMed:19131338). Plays a critical role in maintaining genomic stability and cellular integrity following UV-induced genotoxic stress (PubMed:23398316). Regulates the circadian expression of the core clock components NR1D1 and ARNTL/BMAL1 (PubMed:23398316). Enhances the transcriptional repressor activity of NR1D1 through stabilization of NR1D1 protein levels by preventing its ubiquitination and subsequent degradation (PubMed:23398316). Represses the ligand-dependent transcriptional activation function of ESR2 (PubMed:20074560). Acts as a regulator of PCK1 expression and gluconeogenesis by a mechanism that involves, at least in part, both NR1D1 and SIRT1 (PubMed:24415752). Negatively regulates the deacetylase activity of HDAC3 and can alter its subcellular localization (PubMed:21030595). Positively regulates the beta-catenin pathway (canonical Wnt signaling pathway) and is required for MCC-mediated repression of the beta-catenin pathway (PubMed:24824780). Represses ligand-dependent transcriptional activation function of NR1H2 and NR1H3 and inhibits the interaction of SIRT1 with NR1H3 (PubMed:25661920). Plays an important role in tumor suppression through p53/TP53 regulation; stabilizes p53/TP53 by affecting its interaction with ubiquitin ligase MDM2 (PubMed:25732823). Represses the transcriptional activator activity of BRCA1 (PubMed:20160719). Inhibits SIRT1 in a CHEK2 and PSEM3-dependent manner and inhibits the activity of CHEK2 in vitro (PubMed:25361978). CCAR2_HUMAN,Q8N163 |
| 产品编号 | 产品名称 | 包装 |
| L24979 | CCAR2 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 |
