pLenti-RPA3-sgRNA (RPA3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，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 |
| L30545 | pLenti-RPA3-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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