pLenti-HUNK-sgRNA (HUNK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HUNK基因敲除的质粒(L27815 pLenti-HUNK-sgRNA)、慢病毒(L27816 HUNK Knockout Lentivirus)、HEK293T细胞(L27817 HUNK Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L27818 HUNK Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L27819 HUNK 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。
HUNK基因的基本信息如下：
| Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
| Human | HUNK | 30811 | - | NM_014586 |
| About the gene | |
| Official Symbol | HUNK |
| Previous Symbol | - |
| Official Full Name | hormonally up-regulated Neu-associated kinase |
| Synonyms | - |
| Location | 21q22.11 |
| Gene Type | protein_coding |
| Uniprot ID | P57058 |
| Pathway/Library | others |
| Gene Summary | Hormonally Up-regulated Neu-associated Kinase (HUNK) is a serine/threonine kinase that is a member of the AMP-activated protein kinase (AMPK) family, and was reported to be up-regulated in aggressive subsets of human cancers including breast, ovarian, and colon cancers. Experimental studies indicate a relationship between HUNK and breast cancer progression. Wertheim et al. showed that Hunk wild-type (Hunk+/+) mice had significantly increased lung metastases compared with Hunk-knockout (Hunk−/−) mice when bred into an MMTV-myc background. In addition, tumors derived from Hunk−/− cells retrovirally transduced with a wild-type version of HUNK (HUNK WT) had significantly increased lung metastases compared with those transduced with either a kinase dead version of HUNK (K91M) or a control vector. These data strongly suggest that HUNK promotes breast cancer metastasis, and that this process is dependent on HUNK kinase activity. However, another study provided evidence that HUNK suppresses basal breast cancer metastasis. Quintela-Fandino et al. found that when HUNK was exogenously introduced into basal breast cancer cells, cell motility and tumor metastases were decreased. HUNK directly phosphorylates EGFR in its juxtamembrane domain at threonine 654 (T654), resulting in receptor stabilization and signaling. This effect corresponds to an increase in epithelial to mesenchymal (EMT), cell migration, and invasion in vitro. HUNK stabilizes EGFR expression in primary tumors, which correlates with increased levels of phosphorylation of EGFR at T654, resulting in an increase in lung metastasis in vivo. |
| 产品编号 | 产品名称 | 包装 |
| L27815 | pLenti-HUNK-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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