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plasmids expressing cas9 pdd162  (Addgene inc)


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    Structured Review

    Addgene inc plasmids expressing cas9 pdd162
    Plasmids Expressing Cas9 Pdd162, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 259 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cas9+expression+plasmid/pDD162+(Peft-3%3A%3ACas9+%2B+Empty+sgRNA)+(Plasmid+%2347549)/pm41896213-178-26-31
    Average 95 stars, based on 259 article reviews
    plasmids expressing cas9 pdd162 - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Construct:

    Article Title: Enhanced production of sabinene by engineered Saccharomyces cerevisiae from corn hydrolysates
    Article Snippet: recently, advanced biofuels [1–3].. IPP and DMAPP, derived from both the mevalonate (MVA) pathway and methylerythritol phosphate (MEP) pathway, are condensed to geranyl diphosphate (GPP, precursor to monoterpene) by GPP synthase, which is further converted to monoterpene by various monoterpene synthases [4, 5].. Sabinene is a type of bicyclic monoterpene found in a variety of essential oils of plants, including Salvia officinalis and Origanum species, and is widely used in culinary spices, perfume additives and fine chemicals [6–9].

    Expressing:

    Article Title: Enhanced production of sabinene by engineered Saccharomyces cerevisiae from corn hydrolysates
    Article Snippet: recently, advanced biofuels [1–3].. IPP and DMAPP, derived from both the mevalonate (MVA) pathway and methylerythritol phosphate (MEP) pathway, are condensed to geranyl diphosphate (GPP, precursor to monoterpene) by GPP synthase, which is further converted to monoterpene by various monoterpene synthases [4, 5].. Sabinene is a type of bicyclic monoterpene found in a variety of essential oils of plants, including Salvia officinalis and Origanum species, and is widely used in culinary spices, perfume additives and fine chemicals [6–9].

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. ..

    Article Title: Development of Potent and Selective RIPK1 Degraders Targeting Its Non-Enzymatic Function for Cancer Treatment
    Article Snippet: .. 26 Guide RNA oligos for RIPK1 (foward: CACCGAGCCCATCCAGGGTTAGTTC. reverse: AAACGAACTAACCCTGGATGGGCTC) were cloned into Cas9 expression plasmid (pSpCas9–2A-GFP purchased from Addgene), and co-transfected with donor DNA oligo using FuGENE ® HD Transfection Reagent (Promega). .. GFP positive cells were FACS sorted and single clones were screened for HiBiT signal by Nano Glo ® HiBiT Lytic Detection System (Promega).

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. .. The mScarlet fluorescent reporter was inserted into the EcoRV (New England Biolabs, United States; 10,000 U/mL) site of pCas9 generate pCas9-mScarlet plasmid, allowing visual selection of transformants.

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti-PD-1 therapy.
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma- Aldrich, SCC148) was first transduced with Cas9- expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9- expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Article Title: Mucin-type O -glycans regulate proteoglycan stability and chondrocyte maturation
    Article Snippet: For routine passaging, all cells listed above were detached using 0.05% trypsin-EDTA (Thermo), sub-cultured every 3 to 4 days, and revived from liquid nitrogen after ≤ 10 passages. .. HEK293T cells were co-transfected with Fugene 6 (Promega) and 4 μg each of a viral envelope plasmid (pMD2.g, a gift from Didier Trono and purchased from Addgene, #12259), packaging plasmid (psPAX2, a gift from Didier Trono and purchased from Addgene, #12260), and Cas9 expression plasmid (lentiCas9-Blast, a gift from Feng Zhang and purchased from Addgene, #52962) to generate Cas9 lentiviral particles. ..

    Plasmid Preparation:

    Article Title: Enhanced production of sabinene by engineered Saccharomyces cerevisiae from corn hydrolysates
    Article Snippet: recently, advanced biofuels [1–3].. IPP and DMAPP, derived from both the mevalonate (MVA) pathway and methylerythritol phosphate (MEP) pathway, are condensed to geranyl diphosphate (GPP, precursor to monoterpene) by GPP synthase, which is further converted to monoterpene by various monoterpene synthases [4, 5].. Sabinene is a type of bicyclic monoterpene found in a variety of essential oils of plants, including Salvia officinalis and Origanum species, and is widely used in culinary spices, perfume additives and fine chemicals [6–9].

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. ..

    Article Title: Development of Potent and Selective RIPK1 Degraders Targeting Its Non-Enzymatic Function for Cancer Treatment
    Article Snippet: .. 26 Guide RNA oligos for RIPK1 (foward: CACCGAGCCCATCCAGGGTTAGTTC. reverse: AAACGAACTAACCCTGGATGGGCTC) were cloned into Cas9 expression plasmid (pSpCas9–2A-GFP purchased from Addgene), and co-transfected with donor DNA oligo using FuGENE ® HD Transfection Reagent (Promega). .. GFP positive cells were FACS sorted and single clones were screened for HiBiT signal by Nano Glo ® HiBiT Lytic Detection System (Promega).

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. .. The mScarlet fluorescent reporter was inserted into the EcoRV (New England Biolabs, United States; 10,000 U/mL) site of pCas9 generate pCas9-mScarlet plasmid, allowing visual selection of transformants.

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti-PD-1 therapy.
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma- Aldrich, SCC148) was first transduced with Cas9- expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9- expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Article Title: Mucin-type O -glycans regulate proteoglycan stability and chondrocyte maturation
    Article Snippet: For routine passaging, all cells listed above were detached using 0.05% trypsin-EDTA (Thermo), sub-cultured every 3 to 4 days, and revived from liquid nitrogen after ≤ 10 passages. .. HEK293T cells were co-transfected with Fugene 6 (Promega) and 4 μg each of a viral envelope plasmid (pMD2.g, a gift from Didier Trono and purchased from Addgene, #12259), packaging plasmid (psPAX2, a gift from Didier Trono and purchased from Addgene, #12260), and Cas9 expression plasmid (lentiCas9-Blast, a gift from Feng Zhang and purchased from Addgene, #52962) to generate Cas9 lentiviral particles. ..

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti–PD-1 therapy
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma-Aldrich, SCC148) was first transduced with Cas9-expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9-expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Transformation Assay:

    Article Title: Enhanced production of sabinene by engineered Saccharomyces cerevisiae from corn hydrolysates
    Article Snippet: recently, advanced biofuels [1–3].. IPP and DMAPP, derived from both the mevalonate (MVA) pathway and methylerythritol phosphate (MEP) pathway, are condensed to geranyl diphosphate (GPP, precursor to monoterpene) by GPP synthase, which is further converted to monoterpene by various monoterpene synthases [4, 5].. Sabinene is a type of bicyclic monoterpene found in a variety of essential oils of plants, including Salvia officinalis and Origanum species, and is widely used in culinary spices, perfume additives and fine chemicals [6–9].

    Derivative Assay:

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. ..

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. .. The mScarlet fluorescent reporter was inserted into the EcoRV (New England Biolabs, United States; 10,000 U/mL) site of pCas9 generate pCas9-mScarlet plasmid, allowing visual selection of transformants.

    Mutagenesis:

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. ..

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. .. The mScarlet fluorescent reporter was inserted into the EcoRV (New England Biolabs, United States; 10,000 U/mL) site of pCas9 generate pCas9-mScarlet plasmid, allowing visual selection of transformants.

    Introduce:

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. ..

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. .. The mScarlet fluorescent reporter was inserted into the EcoRV (New England Biolabs, United States; 10,000 U/mL) site of pCas9 generate pCas9-mScarlet plasmid, allowing visual selection of transformants.

    Activity Assay:

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. ..

    Article Title: A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in Mycobacterium abscessus
    Article Snippet: .. The Cas9 expression plasmid was derived from pLJR962 (Addgene #115162) via site-directed mutagenesis to introduce A9D and A599H mutations, restoring nuclease activity and generating pCas9. .. The mScarlet fluorescent reporter was inserted into the EcoRV (New England Biolabs, United States; 10,000 U/mL) site of pCas9 generate pCas9-mScarlet plasmid, allowing visual selection of transformants.

    Clone Assay:

    Article Title: Development of Potent and Selective RIPK1 Degraders Targeting Its Non-Enzymatic Function for Cancer Treatment
    Article Snippet: .. 26 Guide RNA oligos for RIPK1 (foward: CACCGAGCCCATCCAGGGTTAGTTC. reverse: AAACGAACTAACCCTGGATGGGCTC) were cloned into Cas9 expression plasmid (pSpCas9–2A-GFP purchased from Addgene), and co-transfected with donor DNA oligo using FuGENE ® HD Transfection Reagent (Promega). .. GFP positive cells were FACS sorted and single clones were screened for HiBiT signal by Nano Glo ® HiBiT Lytic Detection System (Promega).

    Transfection:

    Article Title: Development of Potent and Selective RIPK1 Degraders Targeting Its Non-Enzymatic Function for Cancer Treatment
    Article Snippet: .. 26 Guide RNA oligos for RIPK1 (foward: CACCGAGCCCATCCAGGGTTAGTTC. reverse: AAACGAACTAACCCTGGATGGGCTC) were cloned into Cas9 expression plasmid (pSpCas9–2A-GFP purchased from Addgene), and co-transfected with donor DNA oligo using FuGENE ® HD Transfection Reagent (Promega). .. GFP positive cells were FACS sorted and single clones were screened for HiBiT signal by Nano Glo ® HiBiT Lytic Detection System (Promega).

    Transduction:

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti-PD-1 therapy.
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma- Aldrich, SCC148) was first transduced with Cas9- expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9- expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti–PD-1 therapy
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma-Aldrich, SCC148) was first transduced with Cas9-expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9-expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Selection:

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti-PD-1 therapy.
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma- Aldrich, SCC148) was first transduced with Cas9- expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9- expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti–PD-1 therapy
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma-Aldrich, SCC148) was first transduced with Cas9-expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9-expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Stable Transfection:

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti-PD-1 therapy.
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma- Aldrich, SCC148) was first transduced with Cas9- expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9- expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).

    Article Title: Loss of PTDSS1 in tumor cells improves immunogenicity and response to anti–PD-1 therapy
    Article Snippet: .. Mouse bladder cancer cell line MB49 (Sigma-Aldrich, SCC148) was first transduced with Cas9-expression plasmid (Addgene 52962) followed by puromycin selection to make Cas9-expressing MB49 stable cell line. .. Mouse CRISPR knockout library was a gift from M. Bassik (Addgene, 1000000122).



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    Image Search Results


    Generation of KRAS knockout clones. (A) Schematic of the CRISPR/Cas9 strategy used to generate KRAS knockout 8988T and KP4 cell lines by targeting exon 2 of KRAS . (B) Immunoblot of RAS - less mouse embryonic fibroblasts (MEFs) reconstituted with BRAF V600E , KRAS WT , HRAS WT , or NRAS WT , confirming specificity of the anti-KRAS antibody (clone 3B10-2F2). (C) Immunoblots showing absence of KRAS protein in 8988T (K275, K328) and KP4 (K22, K63) KRAS knockout clones and baseline PI3K (pAKT) and MAPK (pERK1/2) levels relative to parental cell lines. Loading control is HSP90. Images are representative of n = 3 biological replicates. (D) Bar graphs show quantified pERK/ERK and pAKT/AKT levels relative to the parental cell line (8988T or KP4) (mean ± SEM of n = 3 biological replicates) of immunoblots in ( C ). p -values of repeated measures one-way ANOVA with Tukey’s post hoc test are shown.

    Journal: bioRxiv

    Article Title: Baseline cellular state dictates the molecular impact of KRAS mutant variants in pancreatic cancer cells

    doi: 10.64898/2026.03.10.710185

    Figure Lengend Snippet: Generation of KRAS knockout clones. (A) Schematic of the CRISPR/Cas9 strategy used to generate KRAS knockout 8988T and KP4 cell lines by targeting exon 2 of KRAS . (B) Immunoblot of RAS - less mouse embryonic fibroblasts (MEFs) reconstituted with BRAF V600E , KRAS WT , HRAS WT , or NRAS WT , confirming specificity of the anti-KRAS antibody (clone 3B10-2F2). (C) Immunoblots showing absence of KRAS protein in 8988T (K275, K328) and KP4 (K22, K63) KRAS knockout clones and baseline PI3K (pAKT) and MAPK (pERK1/2) levels relative to parental cell lines. Loading control is HSP90. Images are representative of n = 3 biological replicates. (D) Bar graphs show quantified pERK/ERK and pAKT/AKT levels relative to the parental cell line (8988T or KP4) (mean ± SEM of n = 3 biological replicates) of immunoblots in ( C ). p -values of repeated measures one-way ANOVA with Tukey’s post hoc test are shown.

    Article Snippet: A single-guide RNA (sgRNA) targeting KRAS exon 2 (sequence: 5’-AATTACTACTTGCTTCCTGT-3’) was cloned into the CRISPR-Cas9 expression vector pSpCas9(BB)-2A-GFP (PX458, Addgene plasmid #48138).

    Techniques: Knock-Out, Clone Assay, CRISPR, Western Blot, Control