PiggyBac Gene Editing HR Targeting Vector (MCS1-5’PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3′ PB TR-MCS2)
Products
Catalog Number | Description | Size | Price | Quantity | Add to Cart | |||
---|---|---|---|---|---|---|---|---|
PBHR100A-1 | piggyBac-HR with GFP+Puro markers and TK selection (MCS1-5'PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3′ PB TR-MCS2) for Gene Editing | 10 µg | $1302 |
|
Overview
Overview
Seamless gene editing
Use the PiggyBac Gene Editing HR Targeting Vector (MCS1-5’PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3′ PB TR-MCS2) to get seamless gene editing—leave no trace of vector sequences behind.
This special HR Donor works with the Excision-only PiggyBac Transposase instead of the Cre-LoxP system. Simply proceed with your CRISPR/Cas9 gene editing as usual—clone your homology arms into MCS1 and MCS2, use dual GFP and puromycin selection to find integrants, and enrich for on-target events using negative thymidine kinase (TK) selection (Figure 1).
Why use an HR targeting vector?
Even though gene knock-outs can result from DSBs caused by Cas9 alone, SBI recommends the use of HR targeting vectors (also called HR donor vectors) for more efficient and precise mutation. HR donors can supply elements for positive or negative selection ensuring easier identification of successful mutation events. In addition, HR donors can include up to 6-8 kb of open reading frame for gene knock-ins or tagging, and, when small mutations are included in either 5’ or 3’ homology arms, can make specific, targeted gene edits.
Choose the right HR Targeting Vector for your project
Catalog # | HR Donor Vector | Features* | Application |
|||
---|---|---|---|---|---|---|
Gene | Gene | Gene Edits | Gene Tagging | |||
HR100PA-1 | MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4 | Basic HR Donor | ||||
HR110PA-1 | MCS1-EF1α-RFP-T2A-Puro-pA-MCS2 | Removable RFP marker and puromycin selection | ||||
HR120PA-1 | GFP-pA-LoxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCSPuro-pA-LoxP-MCS | Tag with GFP fusion Removable RFP marker and puromycin selection | ||||
HR130PA-1 | T2A-GFP-pA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCSA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCS | Co-express GFP with “tagged” gene via T2A Removable RFP marker and puromycin selection | ||||
HR150PA-1 | GFP-T2A-Luc-pA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCS | Tag with GFP fusion and co-express luciferase via T2A Removable RFP marker and puromycin selection | ||||
HR180PA-1 | IRES-GFP-pA-loxP-MCS1-EF1α-RFP-T2A-Puro-pA-LoxP-MCS2 | Co-express GFP with “tagged” gene via IRES Removable RFP marker and puromycin selection | ||||
HR210PA-1 | MCS1-LoxP-EF1α-GFP-T2A-Puro-P2A-hsvTK-pA-LoxP-MCS2 | Removable GFP marker, puromycin selection, and TK selection | ||||
HR220PA-1 | GFP-pA-LoxP-EF1α-RFP-T2A-Hygro-pA-LoxP-MCS | Tag with GFP fusion Removable RFP ,arker and hygromycin Selection | ||||
HR410PA-1 | MCS1-EF1α-GFP-T2A-Puro-pA-MCS2 | Removable GFP marker and puromycin selection | ||||
HR510PA-1 | MCS1-EF1α-RFP-T2A-Hygro-pA-MCS2 | Removable RFP marker and hygromycin selection | ||||
HR700PA-1 | MCS1-EF1α-GFP-T2A-Puro-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable GFP marker and puromycin selection | ||||
HR710PA-1 | MCS1-EF1α-RFP-T2A-Hygro-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable RFP marker and hygromycin selection | ||||
HR720PA-1 | MCS1-EF1α-Blasticidin-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable blasticidin selection | ||||
GE602A-1 | pAAVS1D-PGK-MCS-EF1α-copGFPpuro | First generation AAVS1-targeting HR Donor | ||||
GE603A-1 | pAAVS1D-CMV-RFP-EF1α-copGFPpuro | First generation AAVS1-targeting HR Donor (positive control for GE602A-1) | ||||
GE620A-1 | AAVS1-SA-puro-MCS | Second generation AAVS1-targeting HR Donor Promoterless to knock-in any gene or promoter-gene combination | ||||
GE622A-1 | AAVS1-SA-puro-EF1α-MCS | Second generation AAVS1-targeting HR Donor Constitutive expression of your gene-of-interest | ||||
GE624A-1 | AAVS1-SA-puro-MCS-GFP | Second generation AAVS1-targeting HR Donor Create reporter cell lines | ||||
CAS620A-1 | AAVS1-SA-puro-EF1α-hspCas9 | Knock-in Cas9 to the AAVS1 site | ||||
MCS1-5'PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3' PB TR-MCS2 | Use with the PiggyBac Transposon System Enables seamless gene editing with no residual footprint (i.e. completely remove vector sequences) | |||||
*All HR Target Vectors except PBHR100A-1 contain LoxP sites. Any sequences that are integrated between the two LoxP sites can be removed through transient expression of Cre Recombinase. **The clever design of these HR Donors enables enrichment for on-target integration events. A PGK-hsvTK cassette is included outside of the homology arms. Because of this configuration, on-target integration that results from homologous recombination will not include the PGK-hsvTK cassette—only randomly-integrated off-target events will lead to integration of PGK-hsvTK and resulting TK activity. Therefore, TK selection will negatively select against off-target integrants. Click on any one of these vectors to see a diagram of how the negative selection works. |
* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Hera BioLabs, Inc.
How It Works
How It Works
Getting seamless gene editing with the Excision-only PiggyBac Transposase Genome engineering with CRISPR/Cas9For general guidance on using CRISPR/Cas9 technology for genome engineering, including the design of HR Targeting Vectors, take a look at our CRISPR/Cas9 tutorials as well as the following application notes:
CRISPR/Cas9 Gene Knock-Out Application Note (PDF) »CRISPR/Cas9 Gene Editing Application Note (PDF) »
CRISPR/Cas9 Gene Tagging Application Note (PDF) »
Supporting Data
FAQs
Resources
Related Products
Citations
-
Brouwer, I, de Kort, MAC & Lenstra, TL. (2024) Measuring Transcription Dynamics of Individual Genes Inside Living Cells. Methods in molecular biology (Clifton, N.J.). 2024; 2694:235-265. PM ID: 37824008
-
Matta, SK, et al. (2024) Genome-wide and targeted CRISPR screens identify RNF213 as a mediator of interferon gamma-dependent pathogen restriction in human cells. Proceedings of the National Academy of Sciences of the United States of America. 2024; 121(1):e2315865120. PM ID: 38147552
-
Cho, MG, et al. (2024) MRE11 liberates cGAS from nucleosome sequestration during tumorigenesis. Nature. 2024; 625(7995):585-592. PM ID: 38200309
-
Du, M, et al. (2024) Direct observation of a condensate effect on super-enhancer controlled gene bursting. Cell. 2024; 187(2):331-344.e17. PM ID: 38194964
-
Schmitt, J, et al. (2024) Repurposing an endogenous degradation domain for antibody-mediated disposal of cell-surface proteins. EMBO reports. 2024;. PM ID: 38287192
-
Byrnes, AE, et al. (2024) A fluorescent splice-switching mouse model enables high-throughput, sensitive quantification of antisense oligonucleotide delivery and activity. Cell reports methods. 2024; 4(1):100673. PM ID: 38171361
-
Daiki, K, et al. (2024) Blood Endocan as a Biomarker for Breast Cancer Recurrence. Preprint. 2024;. Link: Preprint
-
Koeppel, J, et al. (2024) Randomizing the human genome by engineering recombination between repeat elements. bioRxiv. 2024;. Link: bioRxiv
-
Kortleve, D, et al. (2024) TCR-engineered T-cells directed against Ropporin-1 constitute a safe and effective treatment for triple-negative breast cancer in near-clinical models. bioRxiv. 2024;. Link: bioRxiv
-
Haakonsen, DL, et al. (2024) Stress response silencing by an E3 ligase mutated in neurodegeneration. Nature. 2024; 626(8000):874-880. PM ID: 38297121
-
Gupta, P, et al. (2024) Development of pathophysiologically relevant models of sickle cell disease and β-thalassemia for therapeutic studies. Nature communications. 2024; 15(1):1794. PM ID: 38413594
-
Company, C, et al. (2024) Logical design of synthetic cis-regulatory DNA for genetic tracing of cell identities and state changes. Nature communications. 2024; 15(1):897. PM ID: 38316783
-
Yang, L, et al. (2024) Uncovering receptor-ligand interactions using a high-avidity CRISPR activation screening platform. Science advances. 2024; 10(7):eadj2445. PM ID: 38354234
-
Kubara, K, et al. (2024) Lymph node macrophages drive innate immune responses to enhance the anti-tumor efficacy of mRNA vaccines. Molecular therapy : the journal of the American Society of Gene Therapy. 2024;. PM ID: 38243602
-
Ng-Blichfeldt, J, et al. (2024) Identification of a core transcriptional program driving the human renal mesenchymal-to-epithelial transition. Developmental Cell. 2024;. Link: Developmental Cell
-
Yang, J, Cook, L & Chen, Z. (2024) Systematic evaluation of retroviral LTRs as cis-regulatory elements in mouse embryos. Cell reports. 2024; 43(3):113775. PM ID: 38381606
-
Taglini, F, et al. (2024) DNMT3B PWWP mutations cause hypermethylation of heterochromatin. EMBO reports. 2024;. PM ID: 38291337
-
Tanase-Nakao, K, et al. (2024) Genotype-Phenotype Correlations in Thirty Japanese Patients with Congenital Hypothyroidism Attributable to TG Defects. The Journal of clinical endocrinology and metabolism. 2024;. PM ID: 38373250
-
Alsouri, S, et al. (2024) Actinin-4 controls survival signaling in B cells by limiting the lateral mobility of B-cell antigen receptors. European journal of immunology. 2024;:e2350774. PM ID: 38299456
-
Ke, X, et al. (2024) Establishment of a novel minigenome system for the identification of drugs targeting Nipah virus replication. The Journal of general virology. 2024; 105(1). PM ID: 38180473
- See More
Products
Catalog Number | Description | Size | Price | Quantity | Add to Cart | |||
---|---|---|---|---|---|---|---|---|
PBHR100A-1 | piggyBac-HR with GFP+Puro markers and TK selection (MCS1-5'PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3′ PB TR-MCS2) for Gene Editing | 10 µg | $1302 |
|
Overview
Overview
Seamless gene editing
Use the PiggyBac Gene Editing HR Targeting Vector (MCS1-5’PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3′ PB TR-MCS2) to get seamless gene editing—leave no trace of vector sequences behind.
This special HR Donor works with the Excision-only PiggyBac Transposase instead of the Cre-LoxP system. Simply proceed with your CRISPR/Cas9 gene editing as usual—clone your homology arms into MCS1 and MCS2, use dual GFP and puromycin selection to find integrants, and enrich for on-target events using negative thymidine kinase (TK) selection (Figure 1).
Why use an HR targeting vector?
Even though gene knock-outs can result from DSBs caused by Cas9 alone, SBI recommends the use of HR targeting vectors (also called HR donor vectors) for more efficient and precise mutation. HR donors can supply elements for positive or negative selection ensuring easier identification of successful mutation events. In addition, HR donors can include up to 6-8 kb of open reading frame for gene knock-ins or tagging, and, when small mutations are included in either 5’ or 3’ homology arms, can make specific, targeted gene edits.
Choose the right HR Targeting Vector for your project
Catalog # | HR Donor Vector | Features* | Application |
|||
---|---|---|---|---|---|---|
Gene | Gene | Gene Edits | Gene Tagging | |||
HR100PA-1 | MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4 | Basic HR Donor | ||||
HR110PA-1 | MCS1-EF1α-RFP-T2A-Puro-pA-MCS2 | Removable RFP marker and puromycin selection | ||||
HR120PA-1 | GFP-pA-LoxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCSPuro-pA-LoxP-MCS | Tag with GFP fusion Removable RFP marker and puromycin selection | ||||
HR130PA-1 | T2A-GFP-pA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCSA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCS | Co-express GFP with “tagged” gene via T2A Removable RFP marker and puromycin selection | ||||
HR150PA-1 | GFP-T2A-Luc-pA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCS | Tag with GFP fusion and co-express luciferase via T2A Removable RFP marker and puromycin selection | ||||
HR180PA-1 | IRES-GFP-pA-loxP-MCS1-EF1α-RFP-T2A-Puro-pA-LoxP-MCS2 | Co-express GFP with “tagged” gene via IRES Removable RFP marker and puromycin selection | ||||
HR210PA-1 | MCS1-LoxP-EF1α-GFP-T2A-Puro-P2A-hsvTK-pA-LoxP-MCS2 | Removable GFP marker, puromycin selection, and TK selection | ||||
HR220PA-1 | GFP-pA-LoxP-EF1α-RFP-T2A-Hygro-pA-LoxP-MCS | Tag with GFP fusion Removable RFP ,arker and hygromycin Selection | ||||
HR410PA-1 | MCS1-EF1α-GFP-T2A-Puro-pA-MCS2 | Removable GFP marker and puromycin selection | ||||
HR510PA-1 | MCS1-EF1α-RFP-T2A-Hygro-pA-MCS2 | Removable RFP marker and hygromycin selection | ||||
HR700PA-1 | MCS1-EF1α-GFP-T2A-Puro-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable GFP marker and puromycin selection | ||||
HR710PA-1 | MCS1-EF1α-RFP-T2A-Hygro-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable RFP marker and hygromycin selection | ||||
HR720PA-1 | MCS1-EF1α-Blasticidin-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable blasticidin selection | ||||
GE602A-1 | pAAVS1D-PGK-MCS-EF1α-copGFPpuro | First generation AAVS1-targeting HR Donor | ||||
GE603A-1 | pAAVS1D-CMV-RFP-EF1α-copGFPpuro | First generation AAVS1-targeting HR Donor (positive control for GE602A-1) | ||||
GE620A-1 | AAVS1-SA-puro-MCS | Second generation AAVS1-targeting HR Donor Promoterless to knock-in any gene or promoter-gene combination | ||||
GE622A-1 | AAVS1-SA-puro-EF1α-MCS | Second generation AAVS1-targeting HR Donor Constitutive expression of your gene-of-interest | ||||
GE624A-1 | AAVS1-SA-puro-MCS-GFP | Second generation AAVS1-targeting HR Donor Create reporter cell lines | ||||
CAS620A-1 | AAVS1-SA-puro-EF1α-hspCas9 | Knock-in Cas9 to the AAVS1 site | ||||
MCS1-5'PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3' PB TR-MCS2 | Use with the PiggyBac Transposon System Enables seamless gene editing with no residual footprint (i.e. completely remove vector sequences) | |||||
*All HR Target Vectors except PBHR100A-1 contain LoxP sites. Any sequences that are integrated between the two LoxP sites can be removed through transient expression of Cre Recombinase. **The clever design of these HR Donors enables enrichment for on-target integration events. A PGK-hsvTK cassette is included outside of the homology arms. Because of this configuration, on-target integration that results from homologous recombination will not include the PGK-hsvTK cassette—only randomly-integrated off-target events will lead to integration of PGK-hsvTK and resulting TK activity. Therefore, TK selection will negatively select against off-target integrants. Click on any one of these vectors to see a diagram of how the negative selection works. |
* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Hera BioLabs, Inc.
How It Works
How It Works
Getting seamless gene editing with the Excision-only PiggyBac Transposase Genome engineering with CRISPR/Cas9For general guidance on using CRISPR/Cas9 technology for genome engineering, including the design of HR Targeting Vectors, take a look at our CRISPR/Cas9 tutorials as well as the following application notes:
CRISPR/Cas9 Gene Knock-Out Application Note (PDF) »CRISPR/Cas9 Gene Editing Application Note (PDF) »
CRISPR/Cas9 Gene Tagging Application Note (PDF) »
Supporting Data
FAQs
Citations
-
Brouwer, I, de Kort, MAC & Lenstra, TL. (2024) Measuring Transcription Dynamics of Individual Genes Inside Living Cells. Methods in molecular biology (Clifton, N.J.). 2024; 2694:235-265. PM ID: 37824008
-
Matta, SK, et al. (2024) Genome-wide and targeted CRISPR screens identify RNF213 as a mediator of interferon gamma-dependent pathogen restriction in human cells. Proceedings of the National Academy of Sciences of the United States of America. 2024; 121(1):e2315865120. PM ID: 38147552
-
Cho, MG, et al. (2024) MRE11 liberates cGAS from nucleosome sequestration during tumorigenesis. Nature. 2024; 625(7995):585-592. PM ID: 38200309
-
Du, M, et al. (2024) Direct observation of a condensate effect on super-enhancer controlled gene bursting. Cell. 2024; 187(2):331-344.e17. PM ID: 38194964
-
Schmitt, J, et al. (2024) Repurposing an endogenous degradation domain for antibody-mediated disposal of cell-surface proteins. EMBO reports. 2024;. PM ID: 38287192
-
Byrnes, AE, et al. (2024) A fluorescent splice-switching mouse model enables high-throughput, sensitive quantification of antisense oligonucleotide delivery and activity. Cell reports methods. 2024; 4(1):100673. PM ID: 38171361
-
Daiki, K, et al. (2024) Blood Endocan as a Biomarker for Breast Cancer Recurrence. Preprint. 2024;. Link: Preprint
-
Koeppel, J, et al. (2024) Randomizing the human genome by engineering recombination between repeat elements. bioRxiv. 2024;. Link: bioRxiv
-
Kortleve, D, et al. (2024) TCR-engineered T-cells directed against Ropporin-1 constitute a safe and effective treatment for triple-negative breast cancer in near-clinical models. bioRxiv. 2024;. Link: bioRxiv
-
Haakonsen, DL, et al. (2024) Stress response silencing by an E3 ligase mutated in neurodegeneration. Nature. 2024; 626(8000):874-880. PM ID: 38297121
-
Gupta, P, et al. (2024) Development of pathophysiologically relevant models of sickle cell disease and β-thalassemia for therapeutic studies. Nature communications. 2024; 15(1):1794. PM ID: 38413594
-
Company, C, et al. (2024) Logical design of synthetic cis-regulatory DNA for genetic tracing of cell identities and state changes. Nature communications. 2024; 15(1):897. PM ID: 38316783
-
Yang, L, et al. (2024) Uncovering receptor-ligand interactions using a high-avidity CRISPR activation screening platform. Science advances. 2024; 10(7):eadj2445. PM ID: 38354234
-
Kubara, K, et al. (2024) Lymph node macrophages drive innate immune responses to enhance the anti-tumor efficacy of mRNA vaccines. Molecular therapy : the journal of the American Society of Gene Therapy. 2024;. PM ID: 38243602
-
Ng-Blichfeldt, J, et al. (2024) Identification of a core transcriptional program driving the human renal mesenchymal-to-epithelial transition. Developmental Cell. 2024;. Link: Developmental Cell
-
Yang, J, Cook, L & Chen, Z. (2024) Systematic evaluation of retroviral LTRs as cis-regulatory elements in mouse embryos. Cell reports. 2024; 43(3):113775. PM ID: 38381606
-
Taglini, F, et al. (2024) DNMT3B PWWP mutations cause hypermethylation of heterochromatin. EMBO reports. 2024;. PM ID: 38291337
-
Tanase-Nakao, K, et al. (2024) Genotype-Phenotype Correlations in Thirty Japanese Patients with Congenital Hypothyroidism Attributable to TG Defects. The Journal of clinical endocrinology and metabolism. 2024;. PM ID: 38373250
-
Alsouri, S, et al. (2024) Actinin-4 controls survival signaling in B cells by limiting the lateral mobility of B-cell antigen receptors. European journal of immunology. 2024;:e2350774. PM ID: 38299456
-
Ke, X, et al. (2024) Establishment of a novel minigenome system for the identification of drugs targeting Nipah virus replication. The Journal of general virology. 2024; 105(1). PM ID: 38180473
- See More