BioVector? pCCL.sin.cPPT.EF1α.WPRE Plasmid第三代自失活（SIN）慢病毒轉(zhuǎn)移包裝質(zhì)粒

1. Vector Overview

• Full Name: BioVector? pCCL.sin.cPPT.EF1α.WPRE

• Type: Third-generation, self-inactivating (SIN) lentiviral transfer plasmid.

• Parental Backbone: Derived from the pCCL family of vectors, developed for gene therapy and stable gene transfer.

• Key Application: Used to package lentiviral particles for efficient delivery and stable expression of transgenes in mammalian cells (including dividing and non-dividing cells).

2. Genetic Elements and Features
The plasmid contains several critical functional elements essential for lentiviral production and gene expression:

• 5' LTR (Long Terminal Repeat): Contains the HIV-1 LTR sequence. During packaging, it is replaced by a chimeric promoter (RSV/U3) in the packaging plasmid to initiate transcription of the viral RNA genome.

• Ψ (Psi) Packaging Signal: The primary sequence required for the viral genomic RNA to be packaged into the lentiviral capsid.

• cPPT (central Polypurine Tract): Enhances nuclear import of the viral pre-integration complex, significantly improving transduction efficiency in non-dividing cells.

• EF1α Promoter: Drives the expression of the gene of interest. The human Elongation Factor 1 alpha (EF1α) promoter provides strong and stable expression in a wide range of cell types.

• WPRE (Woodchuck Hepatitis Virus Post-transcriptional Regulatory Element): Enhances transgene expression by increasing RNA stability and facilitating nuclear export of transcripts.

• 3' SIN LTR (Self-Inactivating LTR): Contains a deletion in the U3 region. After reverse transcription and integration into the host genome, this deletion is copied to the 5' LTR, inactivating promoter activity in the provirus to enhance biosafety and prevent mobilization.

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3. Mechanism and Applications

• Lentiviral Production: When co-transfected into packaging cells (e.g., HEK293T) with helper plasmids (packaging and envelope), this vector generates replication-incompetent lentiviral particles.

• Transduction: These particles can transduce target cells. The cPPT facilitates nuclear import, and the EF1α promoter ensures sustained transgene expression.

• Safety: The SIN design reduces the risk of insertional activation of oncogenes.

質(zhì)粒：BioVector? pCCL.sin.cPPT.EF1α.WPRE

1. 載體概述

• 全稱： BioVector? pCCL.sin.cPPT.EF1α.WPRE

• 類型： 第三代自失活（SIN）慢病毒轉(zhuǎn)移質(zhì)粒。

• 親本骨架： 源于pCCL載體家族，專為基因治療和穩(wěn)定基因轉(zhuǎn)移而開發(fā)。

• 主要應(yīng)用： 用于包裝慢病毒顆粒，以在哺乳動(dòng)物細(xì)胞（包括分裂細(xì)胞和非分裂細(xì)胞）中高效遞送和穩(wěn)定表達(dá)目標(biāo)基因。

2. 基因元件與特征
該質(zhì)粒包含多個(gè)對(duì)慢病毒生產(chǎn)和基因表達(dá)至關(guān)重要的功能元件：

• 5' LTR (長(zhǎng)末端重復(fù)序列)： 包含HIV-1 LTR序列。在包裝過程中，該序列會(huì)被包裝質(zhì)粒中的嵌合啟動(dòng)子（RSV/U3）替代，以啟動(dòng)病毒RNA基因組的轉(zhuǎn)錄。

• Ψ (Psi) 包裝信號(hào)： 病毒基因組RNA被包裝入慢病毒衣殼所需的主要序列。

• cPPT (中心多嘌呤束)： 增強(qiáng)病毒預(yù)整合復(fù)合物的核輸入，顯著提高對(duì)非分裂細(xì)胞的轉(zhuǎn)導(dǎo)效率。

• EF1α 啟動(dòng)子： 驅(qū)動(dòng)目標(biāo)基因的表達(dá)。人源延伸因子1 alpha (EF1α) 啟動(dòng)子在多種細(xì)胞類型中提供強(qiáng)健且穩(wěn)定的表達(dá)。

• WPRE (土撥鼠肝炎病毒轉(zhuǎn)錄后調(diào)控元件)： 通過增加RNA穩(wěn)定性和促進(jìn)轉(zhuǎn)錄本的核輸出來增強(qiáng)轉(zhuǎn)基因表達(dá)。

• 3' SIN LTR (自失活LTR)： U3區(qū)域存在缺失。在逆轉(zhuǎn)錄并整合到宿主基因組后，該缺失會(huì)被復(fù)制到5' LTR，從而滅活前病毒的啟動(dòng)子活性，增強(qiáng)生物安全性并防止載體 mobilization。

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3. 機(jī)制與應(yīng)用

• 慢病毒生產(chǎn)： 當(dāng)與輔助質(zhì)粒（包裝質(zhì)粒和包膜質(zhì)粒）共轉(zhuǎn)染到包裝細(xì)胞（如HEK293T）中時(shí)，該載體可生成具有復(fù)制缺陷能力的慢病毒顆粒。

• 轉(zhuǎn)導(dǎo)： 這些顆?？梢赞D(zhuǎn)導(dǎo)靶細(xì)胞。cPPT元件促進(jìn)核輸入，EF1α啟動(dòng)子則確保轉(zhuǎn)基因的持續(xù)表達(dá)。

• 安全性： SIN設(shè)計(jì)降低了插入激活致癌基因的風(fēng)險(xiǎn)。

Supplier生產(chǎn)商：

BioVector NTCC質(zhì)粒載體菌株細(xì)胞蛋白抗體基因保藏中心

電話:400-800-2947

工作QQ/微信同號(hào):1843439339

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