RFWD3基因敲除质粒

pLenti-RFWD3-sgRNA (RFWD3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RFWD3基因敲除的质粒(L10155 pLenti-RFWD3-sgRNA)、慢病毒(L10156 RFWD3 Knockout Lentivirus)、HEK293T细胞(L10157 RFWD3 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L10158 RFWD3 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L10159 RFWD3 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。

RFWD3基因的基本信息如下：

Species	Gene Symbol	Gene ID	GenBank Accession	Transcript
Human	RFWD3	55159	BC059371	NM_018124

 

About the gene
Official Symbol	RFWD3
Previous Symbol	-
Official Full Name	ring finger and WD repeat domain 3
Synonyms	FLJ10520; RNF201; FANCW
Location	16q23.1
Gene Type	protein-coding gene
Uniprot ID	Q6PCD5
Pathway/Library	Ubiquitin Ligases Genes Library
Gene Summary	E3 ubiquitin-protein ligase required for the repair of DNA interstrand cross-links (ICL) in response to DNA damage (PubMed:21504906, PubMed:21558276, PubMed:26474068, PubMed:28575657, PubMed:28575658). Plays a key role in RPA-mediated DNA damage signaling and repair (PubMed:21504906, PubMed:21558276, PubMed:26474068, PubMed:28575657, PubMed:28575658, PubMed:28691929). Acts by mediating ubiquitination of the RPA complex (RPA1, RPA2 and RPA3 subunits) and RAD51 at stalled replication forks, leading to remove them from DNA damage sites and promote homologous recombination (PubMed:26474068, PubMed:28575657, PubMed:28575658). Also mediates the ubiquitination of p53/TP53 in the late response to DNA damage, and acts as a positive regulator of p53/TP53 stability, thereby regulating the G1/S DNA damage checkpoint (PubMed:20173098). May act by catalyzing the formation of short polyubiquitin chains on p53/TP53 that are not targeted to the proteasome (PubMed:20173098). In response to ionizing radiation, interacts with MDM2 and enhances p53/TP53 ubiquitination, possibly by restricting MDM2 from extending polyubiquitin chains on ubiquitinated p53/TP53 (PubMed:20173098). RFWD3_HUMAN,Q6PCD5