pentr u6 vector  (Thermo Fisher)


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    Name:
    BLOCK iT U6 RNAi Entry Vector Kit
    Description:
    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 CassetteOnce 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
    Catalog Number:
    k494500
    Price:
    None
    Applications:
    Cloning|Destination Vectors|Gateway Cloning|RNAi|RNAi, Epigenetics & Non-Coding RNA Research|Vector-Based RNAi
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
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    Structured Review

    Thermo Fisher pentr u6 vector
    Reduction in gag synthesis. 293T cells were transfected with shEXTL2.1, shEXTL2.2, shEXTL2.3, shEXTL3.2 or ctrlEXTL expressed from the <t>pENTR/U6</t> vector. A mock transfection consisting of lipofectamine 2000 reagent only served as a control. After 48 h, 5 μCi/ml 35 SO 4 was added and gag synthesis determined over 4 h. Results are the mean±standard deviation of three replicates, and are expressed as the percentage of the mock control. * Significant difference from mock transfection, P
    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 CassetteOnce 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
    https://www.bioz.com/result/pentr u6 vector/product/Thermo Fisher
    Average 99 stars, based on 58 article reviews
    Price from $9.99 to $1999.99
    pentr u6 vector - by Bioz Stars, 2021-01
    99/100 stars

    Images

    1) Product Images from "Gene silencing of EXTL2 and EXTL3 as a substrate deprivation therapy for heparan sulphate storing mucopolysaccharidoses"

    Article Title: Gene silencing of EXTL2 and EXTL3 as a substrate deprivation therapy for heparan sulphate storing mucopolysaccharidoses

    Journal: European Journal of Human Genetics

    doi: 10.1038/ejhg.2009.143

    Reduction in gag synthesis. 293T cells were transfected with shEXTL2.1, shEXTL2.2, shEXTL2.3, shEXTL3.2 or ctrlEXTL expressed from the pENTR/U6 vector. A mock transfection consisting of lipofectamine 2000 reagent only served as a control. After 48 h, 5 μCi/ml 35 SO 4 was added and gag synthesis determined over 4 h. Results are the mean±standard deviation of three replicates, and are expressed as the percentage of the mock control. * Significant difference from mock transfection, P
    Figure Legend Snippet: Reduction in gag synthesis. 293T cells were transfected with shEXTL2.1, shEXTL2.2, shEXTL2.3, shEXTL3.2 or ctrlEXTL expressed from the pENTR/U6 vector. A mock transfection consisting of lipofectamine 2000 reagent only served as a control. After 48 h, 5 μCi/ml 35 SO 4 was added and gag synthesis determined over 4 h. Results are the mean±standard deviation of three replicates, and are expressed as the percentage of the mock control. * Significant difference from mock transfection, P

    Techniques Used: Transfection, Plasmid Preparation, Standard Deviation

    Quantitative knockdown of EXTL2 and EXTL3 . 293T cells were transfected with psiCHECK-2EXTL2 or psiCHECK-2EXTL3 only, or co-transfected with a psiCHECK-2 target gene construct and corresponding shEXTL or ctrlEXTL expressed from the pENTR/U6 vector. Control cells were mock transfected with lipofectamine 2000 reagent only. A DLR assay was conducted 24 h post-transfection, and results expressed as a percentage of psiCHECK-2 target gene expression. Results are the mean±standard deviation of n =10 replicates. * Significant difference from mock transfection P
    Figure Legend Snippet: Quantitative knockdown of EXTL2 and EXTL3 . 293T cells were transfected with psiCHECK-2EXTL2 or psiCHECK-2EXTL3 only, or co-transfected with a psiCHECK-2 target gene construct and corresponding shEXTL or ctrlEXTL expressed from the pENTR/U6 vector. Control cells were mock transfected with lipofectamine 2000 reagent only. A DLR assay was conducted 24 h post-transfection, and results expressed as a percentage of psiCHECK-2 target gene expression. Results are the mean±standard deviation of n =10 replicates. * Significant difference from mock transfection P

    Techniques Used: Transfection, Construct, Plasmid Preparation, Expressing, Standard Deviation

    Related Articles

    Clone Assay:

    Article Title: Multiple stages of evolutionary change in anthrax toxin receptor expression in humans
    Article Snippet: .. A G was added at the 5’ end of primers for use with a U6 promoter, along with restriction sites for cloning. .. Forward and reverse sgRNAs were synthesized separately by IDT and annealed.

    In Vivo:

    Article Title: Dopamine Axon Targeting in the Nucleus Accumbens in Adolescence Requires Netrin-1
    Article Snippet: .. Importantly, the constructs express GFP under the CMV promoter and the shRNA sequence under the U6 promoter, which allows verification of infection and visualization of the injection site for the in vivo experiments. .. The efficiency of knockdown for shRNA constructs was tested in cultured HEK cells by co-expressing a pBKNetrin-1-Flag plasmid containing the mouse Netrin-1 cDNA cloned into a pBK-CMV vector (Stratagene) with a C-terminal Flag tag (gift of Dr. Andreas Püschel to HM Cooper), Flag expression was then evaluated by western blot using an anti-Flag antibody (1:5000; Cat#F1804; Sigma-Aldrich, Saint Louis, MO, United States).

    shRNA:

    Article Title: Lentivirus-mediated RNAi knockdown of VEGFA in RKO colorectal cancer cells decreases tumor formation and growth in vitro and in vivo
    Article Snippet: .. A shRNA-expressing lentivector was then constructed following the manufacturer’s instructions (BLOCK-iT™ U6 RNAi Entry Vector Kit, Invitrogen) and packaged using the Virapower packaging mix (Invitrogen). .. Virus particles were titered using 293FT cells, as described in the Invitrogen Instruction manual for Virapower Lentiviral Expression Systems (Version E).

    Article Title: Expression of long non‐coding RNA ENSG00000226738 (Lnc KLHDC7B) is enriched in the immunomodulatory triple‐negative breast cancer subtype and its alteration promotes cell migration, invasion, and resistance to cell death
    Article Snippet: .. 2.8 Long non‐coding RNA down‐regulation with shRNA Short hairpin RNA were generated using the BLOCK‐iT™ U6 RNAi Entry Vector Kit (Invitrogen) following the manufacturer's instructions. .. Briefly, pairs of cDNA oligos were designed containing four nucleotide overhangs necessary for directional cloning.

    Article Title: Dopamine Axon Targeting in the Nucleus Accumbens in Adolescence Requires Netrin-1
    Article Snippet: .. Importantly, the constructs express GFP under the CMV promoter and the shRNA sequence under the U6 promoter, which allows verification of infection and visualization of the injection site for the in vivo experiments. .. The efficiency of knockdown for shRNA constructs was tested in cultured HEK cells by co-expressing a pBKNetrin-1-Flag plasmid containing the mouse Netrin-1 cDNA cloned into a pBK-CMV vector (Stratagene) with a C-terminal Flag tag (gift of Dr. Andreas Püschel to HM Cooper), Flag expression was then evaluated by western blot using an anti-Flag antibody (1:5000; Cat#F1804; Sigma-Aldrich, Saint Louis, MO, United States).

    Synthesized:

    Article Title: IGFBP-2 directly stimulates osteoblast differentiation
    Article Snippet: .. The oligonucleotides were synthesized by Nucleic Acids Core Facility at UNC, annealed and ligated into BLOCK-iT™ U6 RNAi Entry Vector (Cat# K4945-00, Life Technologies, Grand Island, NY) following manufacturer’s instructions. ..

    Construct:

    Article Title: Lentivirus-mediated RNAi knockdown of VEGFA in RKO colorectal cancer cells decreases tumor formation and growth in vitro and in vivo
    Article Snippet: .. A shRNA-expressing lentivector was then constructed following the manufacturer’s instructions (BLOCK-iT™ U6 RNAi Entry Vector Kit, Invitrogen) and packaged using the Virapower packaging mix (Invitrogen). .. Virus particles were titered using 293FT cells, as described in the Invitrogen Instruction manual for Virapower Lentiviral Expression Systems (Version E).

    Article Title: Dopamine Axon Targeting in the Nucleus Accumbens in Adolescence Requires Netrin-1
    Article Snippet: .. Importantly, the constructs express GFP under the CMV promoter and the shRNA sequence under the U6 promoter, which allows verification of infection and visualization of the injection site for the in vivo experiments. .. The efficiency of knockdown for shRNA constructs was tested in cultured HEK cells by co-expressing a pBKNetrin-1-Flag plasmid containing the mouse Netrin-1 cDNA cloned into a pBK-CMV vector (Stratagene) with a C-terminal Flag tag (gift of Dr. Andreas Püschel to HM Cooper), Flag expression was then evaluated by western blot using an anti-Flag antibody (1:5000; Cat#F1804; Sigma-Aldrich, Saint Louis, MO, United States).

    Blocking Assay:

    Article Title: Lentivirus-mediated RNAi knockdown of VEGFA in RKO colorectal cancer cells decreases tumor formation and growth in vitro and in vivo
    Article Snippet: .. A shRNA-expressing lentivector was then constructed following the manufacturer’s instructions (BLOCK-iT™ U6 RNAi Entry Vector Kit, Invitrogen) and packaged using the Virapower packaging mix (Invitrogen). .. Virus particles were titered using 293FT cells, as described in the Invitrogen Instruction manual for Virapower Lentiviral Expression Systems (Version E).

    Article Title: IGFBP-2 directly stimulates osteoblast differentiation
    Article Snippet: .. The oligonucleotides were synthesized by Nucleic Acids Core Facility at UNC, annealed and ligated into BLOCK-iT™ U6 RNAi Entry Vector (Cat# K4945-00, Life Technologies, Grand Island, NY) following manufacturer’s instructions. ..

    Article Title: Expression of long non‐coding RNA ENSG00000226738 (Lnc KLHDC7B) is enriched in the immunomodulatory triple‐negative breast cancer subtype and its alteration promotes cell migration, invasion, and resistance to cell death
    Article Snippet: .. 2.8 Long non‐coding RNA down‐regulation with shRNA Short hairpin RNA were generated using the BLOCK‐iT™ U6 RNAi Entry Vector Kit (Invitrogen) following the manufacturer's instructions. .. Briefly, pairs of cDNA oligos were designed containing four nucleotide overhangs necessary for directional cloning.

    Generated:

    Article Title: Expression of long non‐coding RNA ENSG00000226738 (Lnc KLHDC7B) is enriched in the immunomodulatory triple‐negative breast cancer subtype and its alteration promotes cell migration, invasion, and resistance to cell death
    Article Snippet: .. 2.8 Long non‐coding RNA down‐regulation with shRNA Short hairpin RNA were generated using the BLOCK‐iT™ U6 RNAi Entry Vector Kit (Invitrogen) following the manufacturer's instructions. .. Briefly, pairs of cDNA oligos were designed containing four nucleotide overhangs necessary for directional cloning.

    other:

    Article Title: HBO1 (KAT7) Does Not Have an Essential Role in Cell Proliferation, DNA Replication, or Histone 4 Acetylation in Human Cells
    Article Snippet: RNA interference knockdown of HBO1 does not affect cell proliferation or cell cycle dynamics.

    Infection:

    Article Title: Dopamine Axon Targeting in the Nucleus Accumbens in Adolescence Requires Netrin-1
    Article Snippet: .. Importantly, the constructs express GFP under the CMV promoter and the shRNA sequence under the U6 promoter, which allows verification of infection and visualization of the injection site for the in vivo experiments. .. The efficiency of knockdown for shRNA constructs was tested in cultured HEK cells by co-expressing a pBKNetrin-1-Flag plasmid containing the mouse Netrin-1 cDNA cloned into a pBK-CMV vector (Stratagene) with a C-terminal Flag tag (gift of Dr. Andreas Püschel to HM Cooper), Flag expression was then evaluated by western blot using an anti-Flag antibody (1:5000; Cat#F1804; Sigma-Aldrich, Saint Louis, MO, United States).

    Expressing:

    Article Title: Bta-miR-34b regulates milk fat biosynthesis by targeting mRNA decapping enzyme 1A (DCP1A) in cultured bovine mammary epithelial cells
    Article Snippet: .. The gene UXT was used as the endogenous control for mRNA expression analysis , and the gene U6 was used as the endogenous control gene ( ) for miRNA expression analysis. .. Real-time PCR primers for amplification of mRNA ( ) and miRNA ( ) were designed using Primer Premier 5.0 (PREMIER Biosoft International, Inc., Palo Alto, CA) and synthesized by TSINGKE Biological Technology (Xi’an, P.R.

    Sequencing:

    Article Title: Dopamine Axon Targeting in the Nucleus Accumbens in Adolescence Requires Netrin-1
    Article Snippet: .. Importantly, the constructs express GFP under the CMV promoter and the shRNA sequence under the U6 promoter, which allows verification of infection and visualization of the injection site for the in vivo experiments. .. The efficiency of knockdown for shRNA constructs was tested in cultured HEK cells by co-expressing a pBKNetrin-1-Flag plasmid containing the mouse Netrin-1 cDNA cloned into a pBK-CMV vector (Stratagene) with a C-terminal Flag tag (gift of Dr. Andreas Püschel to HM Cooper), Flag expression was then evaluated by western blot using an anti-Flag antibody (1:5000; Cat#F1804; Sigma-Aldrich, Saint Louis, MO, United States).

    Article Title: Neurotransmitter Switching Regulated by miRNAs Controls Changes in Social Preference
    Article Snippet: .. For reverse transcription, sequence-specific primers were added for each miR being tested and for the housekeeping gene U6. .. The assays used (assay number in brackets) were: miR-92b (007028_mat); miR-125a (007071_mat); miR-183 (000484); miR-148a (000470); miR-30d (000420); miR-30c (000419); miR-205a (000509); miR-375 (007201_mat); miR-200b (006005); miR-429 (001077); miR-129 (000590); miR-203 (004590_mat); miR-214 (000517); miR-218 (000521); miR-30e (002223); miR-101 (000438); miR-1b (008022); miR-499 (005772);.

    Injection:

    Article Title: Dopamine Axon Targeting in the Nucleus Accumbens in Adolescence Requires Netrin-1
    Article Snippet: .. Importantly, the constructs express GFP under the CMV promoter and the shRNA sequence under the U6 promoter, which allows verification of infection and visualization of the injection site for the in vivo experiments. .. The efficiency of knockdown for shRNA constructs was tested in cultured HEK cells by co-expressing a pBKNetrin-1-Flag plasmid containing the mouse Netrin-1 cDNA cloned into a pBK-CMV vector (Stratagene) with a C-terminal Flag tag (gift of Dr. Andreas Püschel to HM Cooper), Flag expression was then evaluated by western blot using an anti-Flag antibody (1:5000; Cat#F1804; Sigma-Aldrich, Saint Louis, MO, United States).

    Plasmid Preparation:

    Article Title: Lentivirus-mediated RNAi knockdown of VEGFA in RKO colorectal cancer cells decreases tumor formation and growth in vitro and in vivo
    Article Snippet: .. A shRNA-expressing lentivector was then constructed following the manufacturer’s instructions (BLOCK-iT™ U6 RNAi Entry Vector Kit, Invitrogen) and packaged using the Virapower packaging mix (Invitrogen). .. Virus particles were titered using 293FT cells, as described in the Invitrogen Instruction manual for Virapower Lentiviral Expression Systems (Version E).

    Article Title: IGFBP-2 directly stimulates osteoblast differentiation
    Article Snippet: .. The oligonucleotides were synthesized by Nucleic Acids Core Facility at UNC, annealed and ligated into BLOCK-iT™ U6 RNAi Entry Vector (Cat# K4945-00, Life Technologies, Grand Island, NY) following manufacturer’s instructions. ..

    Article Title: Expression of long non‐coding RNA ENSG00000226738 (Lnc KLHDC7B) is enriched in the immunomodulatory triple‐negative breast cancer subtype and its alteration promotes cell migration, invasion, and resistance to cell death
    Article Snippet: .. 2.8 Long non‐coding RNA down‐regulation with shRNA Short hairpin RNA were generated using the BLOCK‐iT™ U6 RNAi Entry Vector Kit (Invitrogen) following the manufacturer's instructions. .. Briefly, pairs of cDNA oligos were designed containing four nucleotide overhangs necessary for directional cloning.

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  • 99
    Thermo Fisher pentr u6 vector
    Pentr U6 Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pentr u6 vector/product/Thermo Fisher
    Average 99 stars, based on 58 article reviews
    Price from $9.99 to $1999.99
    pentr u6 vector - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier

    91
    Thermo Fisher pentr u6 scr
    Pentr U6 Scr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pentr u6 scr/product/Thermo Fisher
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    pentr u6 scr - by Bioz Stars, 2021-01
    91/100 stars
      Buy from Supplier

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