RAB44基因敲除质粒

pLenti-RAB44-sgRNA (RAB44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RAB44基因敲除的质粒(L13520 pLenti-RAB44-sgRNA)、慢病毒(L13521 RAB44 Knockout Lentivirus)、HEK293T细胞(L13522 RAB44 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L13523 RAB44 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L13524 RAB44 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。

RAB44基因的基本信息如下：

Species	Gene Symbol	Gene ID	GenBank Accession	Transcript
Human	RAB44	401258	-	NM_001257357

 

About the gene
Official Symbol	RAB44
Previous Symbol	RASD3; RASL13
Official Full Name	RAB44, member RAS oncogene family
Synonyms	dJ431A14.3
Location	6p21.2
Gene Type	protein_coding
Uniprot ID	Q7Z6P3
Pathway/Library	Lung Cancer Growth Related Genes Library
Gene Summary	Rab44 is an atypical Rab GTPase that contains some additional domains such as the EF-hand and coiled-coil domains as well as Rab-GTPase domain. Rab44 was an upregulated protein during osteoclast differentiation. Knockdown of Rab44 by small interfering RNA promotes RANKL-induced osteoclast differentiation of the murine monocytic cell line, RAW-D or of bone marrow-derived macrophages (BMMs). In contrast, overexpression of Rab44 prevents osteoclast differentiation. Rab44 was localized in the Golgi complex and lysosomes, and Rab44 overexpression caused an enlargement of early endosomes. A series of deletion mutant studies of Rab44 showed that the coiled-coil domain and lipidation sites of Rab44 is important for regulation of osteoclast differentiation. Mechanistically, Rab44 affects nuclear factor of activated T-cells c1 (NFATc1) signaling in RANKL-stimulated macrophages. Moreover, Rab44 depletion caused an elevation in intracellular Ca2+ transients upon RANKL stimulation, and particularly regulated lysosomal Ca2+ influx. It was suggested that Rab44 negatively regulates osteoclast differentiation by modulating intracellular Ca2+ levels followed by NFATc1 activation.