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BLOCK-iT™ Pol II miR RNAi Expression Vector Kit with EmGFP (Invitrogen™)

The BLOCK-iT™ Pol II miR RNAi Expression Vector Kit with EmGFP combines the advantages of traditional RNAi vectors (stable expression and the ability to use viral delivery) with capabilities for tissue-specific expression and multiple target knockdown from the same transcript. The pcDNA™6.2-GW/EmGFP-miR vector included in the BLOCK-iT™ Pol II miR RNAi Expression Vector Kit with EmGFP is designed to express artificial miRNAs which are engineered to have 100% homology to your target sequence and will result in target cleavage. This vector includes flanking and loop sequences from an endogenous miRNA which directs the excision of the engineered miRNA from a longer Pol II transcript (pre-miRNA). Using Invitrogen's award-winning BLOCK-iT™ RNAi Designer, over 70% of constructs produce more than 70% knockdown. Pol II expression of engineered miRNAs enables:
o Strong expression from the CMV immediate early promoter, with the option to use tissue-specific or other regulated promoters via MultiSite Gateway® recombination
o Co-cistronic expression of an Emerald GFP (EmGFP) reporter in the pcDNA™6.2-GW/EmGFP-miR vector, resulting in very strong correlation of EmGFP expression with the knockdown activity of your miRNA
o Compatibility with many of Invitrogen's Gateway® destination (DEST) vectors for gene expression; including Lentiviral vectors for stable transduction of dividing, non-dividing and primary cell types, the Flp-In™ system for single-site chromosomal integration, and alternative reporter fusion constructs
o Expression of more than one engineered miRNA on the same transcript, allowing the knockdown of multiple genes simultaneously and the generation of synthetic phenotypes
How it Works
For high levels of expression of your miR RNAi sequence, the pcDNA™6.2-GW/miR vector includes the CMV promoter (Figure 1). Simply input a RefSeq accession number or a nucleotide sequence into the free online BLOCK-iT™ RNAi Designer and the software will design optimized miRNAs that have 100% homology to the target of interest. Clone the miRNA into the vector using a fast ligation protocol and transfect for immediate miRNA expression. Expressed miRNA is processed by the endogenous cellular machinery in the nucleus (including Drosha) and then transported into the cytoplasm where it is further processed by Dicer (Figure 2). The fully processed miRNA is then incorporated into RISC where it functions like an siRNA and results in cleavage of the mRNA target.
For a variety of expression options, the miRNA cassette, which contains EmGFP, miR flanking regions, and an miRNA homologous to the target of interest, can be readily moved into a variety of DEST vectors. This occurs through Gateway® recombination reactions in which the miRNA cassette is transferred into a pDONR™ vector (BP reaction) and then into a DEST vector (LR reaction) of choice .

BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System (Invitrogen™)

The BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System combines
Invitrogen's BLOCK-iT™ Pol II miR RNAi and ViraPower™ Lentiviral technologies
to facilitate creation of a replication-incompetent lentivirus that delivers a
microRNA (miRNA) sequence of interest to dividing or non-dividing mammalian
cells for RNA interference (RNAi) analysis thus broadening the
potential RNAi applications beyond those of other traditional retroviral
systems (Naldini, 1998).

The BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System includes:
o A BLOCK-iT™ Pol II miR RNAi Expression Vector Kit for production of an
expression clone containing a double-stranded oligonucleotide encoding a pre-miRNA sequence for expression in mammalian cells. The BLOCK-iT™ Pol II miR RNAi Expression Vector (pcDNA™6.2-GW/miR) provides a rapid and efficient way to clone ds oligo duplexes encoding a desired miRNA target sequence into a vector containing a Pol II promoter (CMV) for use in RNAi analysis. The BLOCK-iT™ Pol II miR RNAi Expression Vector is specifically designed to
allow expression of miRNA sequences and contain specific miR flanking
sequences that allow proper processing of the miRNA.
o The pDONR™221 vector to transfer the premiRNA
expression cassette into the lentiviral expression plasmid using Gateway® Technology. The Gateway® Technology is a universal cloning method that takes advantage of the site-specific recombination properties of bacteriophage lambda (Landy, 1989) to provide a rapid and highly efficient way to move your DNA sequence of interest into multiple vector systems.
o A pLenti6/V5-DEST destination vector into which the pre-miRNA cassette
from the expression clone is transferred using Gateway® Technology.This expression plasmid contains elements that allow packaging of the construct into virions and the Blasticidin resistance marker for selection of stably transduced cell lines.
o Gateway® BP and LR Clonase® II Enzyme Mixes that facilitate the transfer
of the pre-miRNA expression cassette from the expression vector into the
pLenti6/V5-DEST destination vector.
o Components of the ViraPower™ Lentiviral System for production of a
replication-incompetent lentivirus that stably expresses the miRNA of
interest in both dividing and non-dividing mammalian cells. The ViraPower™ Lentiviral Technology facilitates highly efficient, in vitro or invivo delivery of a target gene or RNA to dividing and non-dividing mammalian cells using a replication-incompetent lentivirus. Based on the lentikat™ systemdeveloped by Cell Genesys (Dull et al., 1998), the ViraPower™ LentiviralTechnology possesses features which enhance its biosafety while allowing highlevel expression in a wider range of cell types than traditional retroviral systems.

BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System with EmGFP (Invitrogen™)

The BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System with EmGFP combines Invitrogen's BLOCK-iT™ Pol II miR RNAi and ViraPower™ Lentiviral technologies to facilitate creation of a replication-incompetent lentivirus that delivers a microRNA (miRNA) sequence of interest to dividing or non-dividing mammalian cells for RNA interference (RNAi) analysis thus broadening the potential RNAi applications beyond those of other traditional retroviral systems (Naldini, 1998). The BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System with EmGFP includes: o A BLOCK-iT™ Pol II miR RNAi Expression Vector Kit for production of an expression clone containing a double-stranded oligonucleotide encoding a pre-miRNA sequence for expression in mammalian cells. The BLOCK-iT™ Pol II miR RNAi Expression Vector (pcDNA™6.2-GW/ EmGFP-miR) provides a rapid and efficient way to clone ds oligo duplexes encoding a desired miRNA target sequence into a vector containing a Pol II promoter (CMV) for use in RNAi analysis. The BLOCK-iT™ Pol II miR RNAi Expression Vector is specifically designed to allow expression of miRNA sequences and contain specific miR flanking sequences that allow proper processing of the miRNA. o Co-cistronic expression of an Emerald GFP (EmGFP) reporter in the pcDNA™6.2-GW/EmGFP-miR vector, resulting in very strong correlation of EmGFP expression with the knockdown activity of your miRNA o The pDONR™221 vector to transfer the premiRNA expression cassette into the lentiviral expression plasmid using Gateway® Technology. The Gateway® Technology is a universal cloning method that takes advantage of the site-specific recombination properties of bacteriophage lambda (Landy, 1989) to provide a rapid and highly efficient way to move your DNA sequence of interest into multiple vector systems. o A pLenti6/V5-DEST destination vector into which the pre-miRNA cassette from the expression clone is transferred using Gateway® Technology.This expression plasmid contains elements that allow packaging of the construct into virions and the Blasticidin resistance marker for selection of stably transduced cell lines. o Gateway® BP and LR Clonase® II Enzyme Mixes that facilitate the transfer of the pre-miRNA expression cassette from the expression vector into the pLenti6/V5-DEST destination vector. o Components of the ViraPower™ Lentiviral System for production of a replication-incompetent lentivirus that stably expresses the miRNA of interest in both dividing and non-dividing mammalian cells. The ViraPower™ Lentiviral Technology facilitates highly efficient, in vitro or invivo delivery of a target gene or RNA to dividing and non-dividing mammalian cells using a replication-incompetent lentivirus. Based on the lentikat™ systemdeveloped by Cell Genesys (Dull et al., 1998), the ViraPower™ LentiviralTechnology possesses features which enhance its biosafety while allowing highlevel expression in a wider range of cell types than traditional retroviral systems.

pMIR-REPORT™ miRNA Expression Reporter Vector System (Invitrogen™)

The Ambion® pMIR-REPORT™ miRNA Expression Reporter Vector System is for the measurement of miRNA expression in cells. It contains two mammalian expression vectors—one for cloning miRNA targets and evaluating miRNA regulation, and the other for the normalization of transfection efficiency.

• Clone miRNA targets and evaluate miRNA regulation
• Screen putative miRNA target sequences
• Includes both reporter and control vectors

The pMIR-REPORT™ miRNA Expression Reporter Vector System provides accurate, quantitative, in-cell measurement of miRNA expression. This validated reporter system contains two mammalian expression vectors. The pMIR-REPORT™ Luciferase miRNA Expression Reporter Vector contains firefly luciferase under the control of a mammalian promoter/terminator system, with an miRNA target cloning region downstream of the luciferase translation sequence. This vector is optimized for cloning miRNA targets and evaluating miRNA regulation. A second vector, pMIR-REPORT™ Beta-galactosidase Reporter Control Vector, is provided for normalizing transfection efficiency.

Applications:
pMIR-REPORT™ Luciferase is designed for cloning and testing putative miRNA binding sites. pMIR-REPORT™ Luciferase can be transfected into mammalian cells to evaluate endogenous miRNA expression, or used to evaluate the up- or down-regulation resulting from the transfection of Pre-miR™ miRNA Molecules or Anti-miR™ miRNA Inhibitor Molecules, respectively.

BLOCK-iT™ HiPerform™ Lentiviral Pol II miR RNAi Expression System with EmGFP (Invitrogen™)

The BLOCK-iT™ Pol II miR RNAi Expression Vector Kits combine the advantages of traditional RNAi vectors (stable expression and the ability to use viral delivery) with capabilities for tissue-specific expression and multiple target knockdown from the same transcript. The BLOCK-iT™ Pol II miR RNAi Expression Vector Kits and the BLOCK-iT™ Lentiviral Pol II miR RNAi Expression Systems are designed to express artificial miRNAs which are engineered to have 100% homology to your target sequence and will result in target cleavage. Using Invitrogen’s award-winning BLOCK-iT™ RNAi Designer, over 70% of constructs produce more than 70% knockdown. The Invitrogen Pol II expression family of vectors include:

–Strong expression from the CMV immediate early promoter, with the option to use EF-1α, tissue-specific promoters, or other promoters via MultiSite Gateway® recombination.
–Compatibility with many of Invitrogen’s Gateway® destination (DEST) vectors for gene expression.

–Ability to control the initiation of the RNAi response with a new BLOCK-iT™ Inducible Pol II miR RNAi Expression Vector Kit w/EmGFP (cat. no. K493900).
–A new destination vector in the BLOCK-iT™ HiPerform™ Lentiviral PolII miR RNAi Expression System with EmGFP. The new vector contains an mRNA stabilizing sequence (WPRE) and a nuclear import sequence (cPPT) which generate up to 5-fold higher virus titers and EmGFP expression levels. Additionally, blasticidin resistance is expressed from the mouse PGK-1 promoter to avoid shut-down after multiple passages.

–Co-cistronic expression of Emerald GFP (EmGFP), resulting in correlation of EmGFP expression with knockdown from your miR RNAi.

–Expression of more than one engineered miR RNAi sequence on the same transcript, allowing the knockdown of multiple genes simultaneously and the generation of synthetic phenotypes

The new BLOCK-iT™ Inducible Pol II miR RNAi Expression Vector Kit with EmGFP provides the ability to regulate RNAi experiments. This kit contains the pT-REx-DEST30 Gateway® vector which after simple cloning and shuttling techniques, produces a miR RNAi expression vector suitable for inducible knockdown. The pT-REx-DEST30 Gateway® vector contains the CMV promoter with two copies of the tetracycline operator (tetO2) sequence allowing high-level and regulated expression. This permits the study of loss of function in a stably transfected cell line even if the gene of interest is essential. Also, induction of miR RNAi expression can be halted so phenotypic changes can be measured during recovery of gene function.

For a variety of expression options, the miR RNAi cassette, which contains EmGFP (pcDNA™6.2-GW/EmGFP-miR vector only), miR flanking regions, and an miRNA homologous to the target of interest, can be readily moved into a variety of DEST vectors. This occurs through Gateway® recombination reactions in which the miR RNAi cassette is transferred into a pDONR™ vector (BP reaction) and then into a DEST vector (LR reaction) of choice. The HiPerform™ system has the new pLenti6.4/R4R2/V5-DEST vector for high virus titers and EmGFP expression.

pAd/BLOCK-iT™-DEST RNAi Gateway Vector (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

BLOCK-iT™ Inducible Pol II miR RNAi Expression Vector Kit with EmGFP (Invitrogen™)

The BLOCK-iT™ Pol II miR RNAi Expression Vector Kits combine the advantages of traditional RNAi vectors (stable expression and the ability to use viral delivery) with capabilities for tissue-specific expression and multiple target knockdown from the same transcript. The BLOCK-iT™ Pol II miR RNAi Expression Vector Kits and the BLOCK-iT™ Lentiviral Pol II miR RNAi Expression Systems are designed to express artificial miRNAs which are engineered to have 100% homology to your target sequence and will result in target cleavage. Using Invitrogen’s award-winning BLOCK-iT™ RNAi Designer, over 70% of constructs produce more than 70% knockdown. The Invitrogen Pol II expression family of vectors include:

–Strong expression from the CMV immediate early promoter, with the option to use tissue-specific or other regulated promoters via MultiSite Gateway® recombination.
–Compatibility with many of Invitrogen’s Gateway® destination (DEST) vectors for gene expression.

–Ability to control the initiation of the RNAi response with a new BLOCK-iT™ Inducible Pol II miR RNAi Expression Vector Kit w/EmGFP.
–A new destination vector in the BLOCK-iT™ HiPerform™ Lentiviral PolII miR RNAi Expression System with EmGFP. The new vector contains an mRNA stabilizing sequence (WPRE) and a nuclear import sequence (cPPT) which generate up to 5-fold higher virus titers and EmGFP expression levels. Additionally, blasticidin resistance is expressed from the mouse Pgk-1 promoter to avoid shut-down after multiple passages.

–Co-cistronic expression of Emerald GFP (EmGFP), resulting in correlation of EmGFP expression with knockdown from your miR RNAi.

–Expression of more than one engineered miR RNAi sequence on the same transcript, allowing the knockdown of multiple genes simultaneously and the generation of synthetic phenotypes

The new BLOCK-iT™ Inducible Pol II miR RNAi Expression Vector Kit with EmGFP provides the ability to regulate RNAi experiments. This kit contains the pT-REx-DEST30 Gateway® vector which after simple cloning and shuttling techniques, produces a miR RNAi expression vector suitable for inducible knockdown. The pT-REx-DEST30 Gateway® vector contains the CMV promoter with two copies of the tetracycline operator (tetO2) sequence allowing high-level and regulated expression. This permits the study of loss of function in a stably transfected cell line even if the gene of interest is essential. Also, induction of miR RNAi expression can be halted so phenotypic changes can be measured during recovery of gene function.

For a variety of expression options, the miR RNAi cassette, which contains EmGFP (pcDNA™6.2-GW/EmGFP-miR vector only), miR flanking regions, and an miRNA homologous to the target of interest, can be readily moved into a variety of DEST vectors. This occurs through Gateway® recombination reactions in which the miR RNAi cassette is transferred into a pDONR™ vector (BP reaction) and then into a DEST vector (LR reaction) of choice. The HiPerform™ system has the new pLenti6.4/RF R2/V5-DEST destination vector for high virus titers and EmGFP expression.

BLOCK-iT™ Adenoviral RNAi Expression System (Invitrogen™)

The pAd/BLOCK-iT™-DEST RNAi Gateway® Vector can be used for the efficient delivery and transient expression of a short hairpin RNA (shRNA) in vivo from an adenoviral vector. A novel cloning process places an ~50-bp DNA oligonucleotide immediately following a U6 pol III promoter into the BLOCK-iT™ U6 entry vector. After efficient recombination of the entry vector into the pAd/BLOCK-iT™-DEST vector, followed by viral production and transduction, the shRNA driven by the U6 promoter can be transiently expressed in most dividing or non-dividing mammalian cell types. The shRNA generated avoids the hosts defense mechanism and will be effective at producing the RNAi gene knockdown response.

The pAd/BLOCK-iT™-DEST vector (Figure 1) offers:

• A promoterless region surrounded by attR sites for efficient recombination with the attL-flanked U6 Gateway® entry vector containing the RNAi cassette or any attL-flanked promoter and gene sequence
• All of the required components for efficient adenoviral packaging and delivery of the shRNA of interest. With this vector, transient analysis of gene knockdown in both dividing and non-dividing mammalian cell types and animal models can be achieved.

BLOCK-iT™ Pol II miR RNAi Expression Vector Kit (Invitrogen™)

The BLOCK-iT™ Pol II miR RNAi Expression Vector Kit combines the advantages of traditional RNAi vectors (stable expression and the ability to use viral delivery) with capabilities for tissue-specific expression and multiple target knockdown from the same transcript. The pcDNA™6.2-GW/miR vector included in the BLOCK-iT™ Pol II miR RNAi Expression Vector Kit is designed to express artificial miRNAs which are engineered to have 100% homology to your target sequence and will result in target cleavage. This vector includes flanking and loop sequences from an endogenous miRNA which directs the excision of the engineered miRNA from a longer Pol II transcript (pre-miRNA). Using Invitrogen's award-winning BLOCK-iT™ RNAi Designer, over 70% of constructs produce more than 70% knockdown. Pol II expression of engineered miRNAs enables:
o Strong expression from the CMV immediate early promoter, with the option to use tissue-specific or other regulated promoters via MultiSite Gateway® recombination
o Compatibility with many of Invitrogen's Gateway® destination (DEST) vectors for gene expression; including Lentiviral vectors for stable transduction of dividing, non-dividing and primary cell types, the Flp-In™ system for single-site chromosomal integration, and alternative reporter fusion constructs
o Expression of more than one engineered miRNA on the same transcript, allowing the knockdown of multiple genes simultaneously and the generation of synthetic phenotypes
How it Works
For high levels of expression of your miR RNAi sequence, the pcDNA™6.2-GW/miR vector includes the CMV promoter (Figure 1). Simply input a RefSeq accession number or a nucleotide sequence into the free online BLOCK-iT™ RNAi Designer and the software will design optimized miRNAs that have 100% homology to the target of interest. Clone the miRNA into the vector using a fast ligation protocol and transfect for immediate miRNA expression. Expressed miRNA is processed by the endogenous cellular machinery in the nucleus (including Drosha) and then transported into the cytoplasm where it is further processed by Dicer (Figure 2). The fully processed miRNA is then incorporated into RISC where it functions like an siRNA and results in cleavage of the mRNA target.
For a variety of expression options, the miRNA cassette, miR flanking regions, and an miRNA homologous to the target of interest, can be readily moved into a variety of DEST vectors. This occurs through Gateway® recombination reactions in which the miRNA cassette is transferred into a pDONR™ vector (BP reaction) and then into a DEST vector (LR reaction) of choice .

BLOCK-iT™ Lentiviral RNAi Zeo Gateway™ Vector Kit (Invitrogen™)

The BLOCK-iT™ Lentiviral RNAi Zeo Gateway® Vector Kit contains the pLenti4/BLOCK-iT™-DEST expression vector which enables lentiviral delivery and genomic integration of DNA coding for shRNA. Once expressed, the shRNA is processed by cellular machinery and initiates target-specific RNAi. The pLenti4/BLOCK-iT™-DEST vector offers:

Gateway® Technology for efficient recombination of the RNAi cassette from the BLOCK-iT™ inducible pENTR™/H1/TO vector

• All required components for efficient lentiviral packaging, delivery, and integration of the shRNA
• Zeocin™ selection marker for fast selection of clonal cell lines containing the RNAi cassette

BLOCK-iT™ U6 RNAi Entry Vector Kit (Invitrogen™)

The BLOCK-iT™ U6 RNAi Entry Vector provides a simple, streamlined approach to cloning short hairpin RNA (shRNA) sequences for testing in transient transfections for RNA interference (RNAi), a preferred technique for analyzing gene downregulation. An easy cloning process places an ~50-bp oligonucleotide DNA immediately following a U6 pol III type promoter (Figure 1). Expression of this RNAi cassette forms a shRNA molecule in the cell that will be processed and act as short interfering RNA (siRNA) that will generate the RNAi effect.

Delivery of the U6 RNAi Cassette
Once cloning is complete, the U6 Entry Vector is ready to be used immediately in transient transfections using a reagent such as Lipofectamine™ 2000. This makes this system ideal for initial shRNA screenings in many mammalian cell types. As an alternative in hard-to-transfect or non-dividing cell types, or to deliver to animal model systems, the RNAi cassette can easily be recombined into a BLOCK-iT™ Viral RNAi vector. For stable shRNA lentiviral delivery and expression, recombine the BLOCK-iT™ U6 Entry Vector with the pLenti6/BLOCK-iT™ RNAi Vector. For transient delivery to challenging cell types using adenoviral transduction, recombine with the pAd/BLOCK-iT™ RNAi Vector.

BLOCK-iT™ Lentiviral RNAi Expression System (Invitrogen™)

The pLenti6/BLOCK-iT™-DEST expression vector provided in the BLOCK-iT™ Lentiviral RNAi Expression System can be used to efficiently introduce and stably express short hairpin RNA (shRNA) in vivo from a lentiviral vector. A novel cloning process places an ~50-bp DNA oligonucleotide immediately following a U6 pol III promoter into the BLOCK-iT™ U6 entry vector. The oligonucleotide is designed to express RNA that forms a stem-loop structure containing the sense and antisense regions of your target gene of interest. This shRNA is then recombined into the pLenti6/BLOCK-iT™-DEST vector. After viral production and transduction, the shRNA driven by the U6 promoter becomes stably integrated as an RNAi cassette. The shRNA generated avoids the hosts defense mechanism and will be effective at producing the RNAi gene knockdown response (Figure 1).

The pLenti6/BLOCK-iT™-DEST vector (Figure 2) offers:

attR sites for efficient recombination with the attL-flanked U6 Gateway® entry vector containing the RNAi cassette
• All of the required components for efficient lentiviral packaging and delivery of the shRNA of interest
• Blasticidin selection marker for fast, efficient selection of stable cell lines expressing the shRNA Using the BLOCK-iT Lentiviral RNAi Expression System, long-term analysis of gene blocking in both dividing and non-dividing mammalian cell types and animal models can be achieved.

The BLOCK-iT™ RNAi U6 Entry Vector Kit allows streamlined cloning of shRNA target sequences for testing in transient experiments. Selected RNAi expression cassettes are quickly and efficiently recombined from the BLOCK-iT™ RNAi U6 entry vector into the pLenti6/BLOCK-iT™-DEST vector via a standard Gateway® LR recombination reaction (Figure 3).

BLOCK-iT™ Lentiviral RNAi Gateway™ Vector Kit (Invitrogen™)

The pLenti6/BLOCK-iT™-DEST expression vector provided in the BLOCK-iT™ Lentiviral RNAi Expression System can be used to efficiently introduce and stably express short hairpin RNA (shRNA) in vivo from a lentiviral vector. A novel cloning process places an ~50-bp DNA oligonucleotide immediately following a U6 pol III promoter into the BLOCK-iT™ U6 entry vector. The oligonucleotide is designed to express RNA that forms a stem-loop structure containing the sense and antisense regions of your target gene of interest. This shRNA is then recombined into the pLenti6/BLOCK-iT™-DEST vector. After viral production and transduction, the shRNA driven by the U6 promoter becomes stably integrated as an RNAi cassette. The shRNA generated avoids the hosts defense mechanism and will be effective at producing the RNAi gene knockdown response (Figure 1).

The pLenti6/BLOCK-iT™-DEST vector (Figure 2) offers:

attR sites for efficient recombination with the attL-flanked U6 Gateway® entry vector containing the RNAi cassette
• All of the required components for efficient lentiviral packaging and delivery of the shRNA of interest
• Blasticidin selection marker for fast, efficient selection of stable cell lines expressing the shRNA Using the BLOCK-iT™ Lentiviral RNAi Expression System, long-term analysis of gene blocking in both dividing and non-dividing mammalian cell types and animal models can be achieved.

The BLOCK-iT™ RNAi U6 Entry Vector Kit allows streamlined cloning of shRNA target sequences for testing in transient experiments. Selected RNAi expression cassettes are quickly and efficiently recombined from the BLOCK-iT™ RNAi U6 entry vector into the pLenti6/BLOCK-iT™-DEST vector via a standard Gateway® LR recombination reaction (Figure 3).