产品编号 | 产品名称 | 产品包装 | 产品价格 |
L30545 | RPA3基因敲除质粒 | 5µg | 999.00元 |
L30546 | RPA3基因敲除慢病毒 | 10^8 TU | 6999.00元 |
L30547 | RPA3基因敲除HEK293T细胞 | 1支/瓶 | 7999.00元 |
L30548 | RPA3基因敲除HEK293T细胞RIPA裂解液 | 100µg | 1258.00元 |
L30549 | RPA3基因敲除HEK293T细胞Trizol裂解液 | 500µl | 1258.00元 |
RPA3 Knockout HEK293T Cells (RPA3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RPA3基因敲除的质粒(L30545 pLenti-RPA3-sgRNA)、慢病毒(L30546 RPA3 Knockout Lentivirus)、HEK293T细胞(L30547 RPA3 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L30548 RPA3 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L30549 RPA3 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。
RPA3基因的基本信息如下:
Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
Human | RPA3 | 6119 | BC005264 | NM_002947 |
About the gene | |
Official Symbol | RPA3 |
Previous Symbol | - |
Official Full Name | replication protein A3 |
Synonyms | REPA3 |
Location | 7p21.3 |
Gene Type | protein-coding gene |
Uniprot ID | P35244 |
Pathway/Library | others |
Gene Summary | As part of the heterotrimeric replication protein A complex (RPA/RP-A), binds and stabilizes single-stranded DNA intermediates that form during DNA replication or upon DNA stress. It prevents their reannealing and in parallel, recruits and activates different proteins and complexes involved in DNA metabolism. Thereby, it plays an essential role both in DNA replication and the cellular response to DNA damage (PubMed:9430682). In the cellular response to DNA damage, the RPA complex controls DNA repair and DNA damage checkpoint activation. Through recruitment of ATRIP activates the ATR kinase a master regulator of the DNA damage response (PubMed:24332808). It is required for the recruitment of the DNA double-strand break repair factors RAD51 and RAD52 to chromatin, in response to DNA damage. Also recruits to sites of DNA damage proteins like XPA and XPG that are involved in nucleotide excision repair and is required for this mechanism of DNA repair (PubMed:7697716). Plays also a role in base excision repair (BER), probably through interaction with UNG (PubMed:9765279). Also recruits SMARCAL1/HARP, which is involved in replication fork restart, to sites of DNA damage. May also play a role in telomere maintenance. RPA3 has its own single-stranded DNA-binding activity and may be responsible for polarity of the binding of the complex to DNA (PubMed:19010961). As part of the alternative replication protein A complex, aRPA, binds single-stranded DNA and probably plays a role in DNA repair. Compared to the RPA2-containing, canonical RPA complex, may not support chromosomal DNA replication and cell cycle progression through S-phase. The aRPA may not promote efficient priming by DNA polymerase alpha but could support DNA synthesis by polymerase delta in presence of PCNA and replication factor C (RFC), the dual incision/excision reaction of nucleotide excision repair and RAD51-dependent strand exchange (PubMed:19996105). RFA3_HUMAN,P35244 |
产品编号 | 产品名称 | 包装 |
L30547 | RPA3 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 |