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Request a custom collection of TrueGuide synthetic gRNA in a convenient arrayed plate format.
TrueGuide Synthetic CRISPR gRNA
Invitrogen TrueGuide Synthetic gRNAs are ready-to-transfect CRISPR guide RNAs designed by a proprietary algorithm for high-efficiency editing. These predesigned, guaranteed guide RNAs offer targeted knockdown with high specificity across the human and mouse genomes. To get started, use the search tool above to search for your gene
Note: TrueGuide crRNA:tracrRNA and unmodified synthetic single guide RNA (sgRNA) have been discontinued. We now only offer our pre-designed modified synthetic sgRNA, which demonstrates superior efficiency across more cell types than the two-part system. Predesigned unmodified sgRNA can still be ordered from the Custom gRNA Order tool. We apologize for any inconvenience this may cause. If you have questions, contact us at techservices@thermofisher.com.
![]() TrueGuide 1-piece modified Synthetic gRNA | |
Predesigned knockouts for mouse and human cells | |
Custom sequences | |
Validated controls available | |
Available in plate format | |
Ready-to-transfect | |
Chemical modifications* | |
Ideal for primary and stem cells |
*Chemical modifications include 2´O-Methyl analogs and phosphorothioate linkages to increase editing efficiency and protect against nuclease degradation.
Lentiviral CRISPR gRNA
Check out our LentiArray CRISPR gRNA if you need to enrich edited cells, are working with difficult-to-transfect cells, or require long-term sgRNA expression.
Cell line-specific transfection conditions using Lipofectamine CRISPRMAX Reagent
The following cell line-specific conditions are provided as a starting point for transfecting wild-type cells with TrueGuide Synthetic gRNA and TrueCut Cas9 Protein v2 using the Lipofectamine CRISPRMAX Transfection Reagent. Further optimization of the transfection conditions may be necessary for best results.
NOTE: Scroll to the right within the table to see all data.
Cell type | Source | Media | Cell seeding density/well (×103) one day before transfection | TrueCut Cas9 Protein v2/gRNA (ng/pmoles) | Lipofectamine Cas9 Plus Reagent/well (μL) | Lipofectamine CRISPRMAX Reagent/well (μL) | ||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Well format | — | — | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well |
HEK293 | Human embryonic kidney | DMEM | 18 | 90 | 450 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.4 | 2 | 10 |
U2OS | Human osteosarcoma | McCoy5A | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
A549 | Human epithelial lung carcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
THP1 | Human peripheral blood monocyte leukemia | RPMI | 10 | 50 | 250 | 400/2.4 | 2000/12 | 10000/60 | 0.8 | 4 | 20 | 0.3 | 1.5 | 7.5 |
K562* | Human leukemia bone marrow | RPMI | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
iPSC* | Human induced pluripotent stem cells | Essential 8 | 8 | 40 | 200 | 300/2 | 1500/10 | 7500/50 | 0.6 | 3 | 15 | 0.3 | 1.5 | 7.5 |
HepG2 | Human hepatocellular carcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
MDA- MB231 | Human epithelial (breast) adenocarcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
N2A | Mouse brain neuroblastoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
*Use the Neon Transfection System for higher editing efficiency. |
Cell-line specific electroporation conditions using Neon system
The following cell line specific conditions are provided as a starting point for transfecting wild-type cells with TrueGuide Synthetic gRNA and TrueCut Cas9 Protein v2 using the Neon Transfection System 10 μL Kit. Further optimization of the electroporation or nucleofection conditions may be necessary for best results.
Cell type | Source | Media | Number of cells/10-μL reaction (× 103) | TrueCut Cas9 Protein v2/gRNA (ng/pmoles) | Neon electroporation conditions* |
---|---|---|---|---|---|
Well format | — | — | 24-well | ||
HEK293 | Human embryonic kidney | DMEM | 150 | 1250/7.5 | 1150 V/20 ms/2 pulses |
U2OS | Human osteosarcoma | McCoy5A | 150 | 1250/7.5 | 1400 V/15 ms/4 pulses |
A549 | Human epithelial lung carcinoma | DMEM | 120 | 1250/7.5 | 1200 V/20 ms/4 pulses |
THP1 | Human peripheral blood monocyte leukemia | RPMI | 200 | 2000/12 | 1700 V/20 ms/1 pulse (#5) |
K562 | Human leukemia bone marrow | RPMI | 200 | 1250/7.5 | 1700 V/20 ms/1 pulse (#5) |
iPSC | Human induced pluripotent stem cells | Essential 8 | 80 | 1500/10 | 1200 V/20 ms/2 pulses (#14) |
iPSC | Human induced pluripotent stem cells | StemFlex | 80 | 1500/10 | 1200 V/30 ms/1 pulse (#7) |
Human primary T-cell | Healthy donor derived | OpTmizer + 2% human serum | 200 | 1250/7.5 | 1600 V/10 ms/3 pulses (#24) |
Jurkat T-cell | Human peripheral blood lymphocyte | RPMI | 200 | 1250/7.5 | 1700 V/20 ms/1 pulse (#5) |
HepG2 | Human hepatocellular carcinoma | DMEM | 120 | 1250/7.5 | 1300 V/30 ms/1 pulse (#8) |
N2A | Mouse brain neuroblastoma | DMEM | 100 | 1250/7.5 | 1400 V/30 ms/1 pulse (#9) |
*Recommendations for the Neon electroporation settings are based on the culture conditions tested. |
Application data
Up to 90% functional knockout in T-cells
Figure 1. High efficiency functional knockout in T-cells. Human T cells were isolated and activated using Invitrogen Dynabeads magnetic beads (Cat. Nos. 11344D and 11141D, respectively) and then transfected with TrueCut Cas9 Protein v2 and TrueGuide Synthetic gRNA for T cell receptor alpha (TRAC) or beta (TRBC) regions using the Invitrogen Neon delivery system. Following transfection, editing efficiency was measured by (A) the Invitrogen GeneArt Cleavage Detection assay, or by (B,C) functional knockout measured by % T cell receptor negative (TCR–) cells using the Invitrogen Attune NxT Flow Cytometer. Cells analyzed by flow cytometry were stained with T cell receptor–specific antibody conjugated to PE. Here we see up to 90% functional knockout in T cells using our optimized protocol.
Genome editing in cancer cells
Figure 2. Prevail with tools to help unravel the complexities of cancer cells. A wide range of cancer cell lines and gene targets were tested to determine editing efficiencies using the modified single TrueGuide Synthetic gRNA (Cat. No. A35511) complexed with TrueCut Cas9 Protein V2 (Cat. No. A36497). A two-fold increase in efficiency was observed as compared to editing systems from competitors. Here the Cas9 RNP complex was delivered into cell lines indicated using Lipofectamine CRISPRMAX (Cat. No. CMAX00003) and at 72 hours post-transfection, editing efficiency was accessed by Ion Torrent NGS sequencing (NGS) or GeneArt Genomic Cleavage Detection (GCD) Kit (Cat. No. A24372).
Editing efficiencies that outperform the competition
Figure 3. Invitrogen CRISPR genome editing tools offer consistently higher editing efficiency in difficult targets, up to 2 times higher than the competition. Genome editing of multiple gene targets was performed with TrueCut Cas9 Protein V2 and corresponding TrueGuide Synthetic gRNAs. Delivery was achieved using recommended transfection protocols and Lipofectamine CRISPRMAX. The graphs also compare the same experiments using products and recommended protocols another manufacturer. With the Invitrogen system, cleavage efficiency (as measured by %Indel) is improved for low efficient loci (PRKCG-T1 and CMPK1-T1) and shows consistently superior efficiency when compared to competitor products and protocols even for challenging loci.
Cell viability
Figure 4. Optimized protocols designed for minimal cell toxicity. This data set illustrates how the combination of gRNAs synthesized using our proprietary design and delivered using our recommended transfection protocols minimize impact on overall cell viability. Lipofectamine CRISPRMAX were used to complex and deliver TrueCut Cas9 Protein V2 and target specific TrueGuide Synthetic gRNAs for editing several loci including two known to be challenging to edit (PRKCG-T1 and CMPK1-T1). Cell viability was measured by PrestoBlue.
User guide
CRISPR validated protocol collection
Frequently asked questions: TrueGuide Synthetic gRNA
Get your CRISPR guide RNA questions answered
gRNA, or guide RNA, is part of the CRISPR-Cas9 system. The gRNA molecule guides the Cas9 protein to a specific genomic locus via base pairing between the crRNA sequence and the target sequence. The CRISPR-Cas9 system is composed of a short noncoding gRNA that has two components: a target-specific CRISPR RNA (crRNA) and an auxiliary trans-activating crRNA (tracrRNA).
sgRNA stands for single guide RNA. As opposed to the native S. pyogenes CRISPR-Cas9 system that uses a two-part guide (CRISPR RNA [crRNA] hybridized with a tracrRNA), an sgRNA combines both RNAs into a single, long RNA. sgRNA are typically 100 nt in length and can be expressed from a vector or chemically synthesized.
crRNA is one strand of the conventional two-part guide RNA system. The TrueGuide crRNA:tracrRNA two-part guide RNA product format has been discontinued to simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
tracrRNA is one strand of the conventional two-part guide RNA system. The TrueGuide crRNA:tracrRNA two-part guide RNA product format has been discontinued to simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
gRNAs are available as chemically synthesized RNA oligos or generated as in vitro transcribed (IVT) gRNAs using the GeneArt Precision gRNA synthesis kit. The IVT gRNA format can also be purchased as a custom ready-to-use option from GEMServices@thermofisher.com. Chemically synthesized gRNA are available from Thermo Fisher Scientific in three different formats:
- Modified sgRNA format: Invitrogen customers can purchase the single guide RNA (sgRNA), which is comprised of both the crRNA and tracrRNA as a single long RNA with 2′ O-Methyl and phosphorothioate modifications on both 5′ and 3′ ends of the molecule. These modifications enhance editing efficiency by increasing binding to the target site and inhibiting nuclease degradation, respectively.
- Two-part format: The TrueGuide crRNA:tracrRNA two-part guide RNA product format has been discontinued to simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
- Unmodified sgRNA format: Customers can purchase a single guide RNA (sgRNA) without any chemical modifications from the Custom gRNA Order tool. The predesigned option for unmodified sgRNA has been discontinued. To simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
Chemical modifications include 2’ O-Methyl analogs and phosphorothioate internucleotide linkages at the 5′ and 3′ ends of the molecule. These modifications enhance editing efficiency by increasing binding to the target site and inhibiting nuclease degradation, respectively.
TrueGuide synthetic gRNA along with TrueCut Cas9 Protein V2 offers maximum editing efficiency across broad cell types while minimizing cell toxicity and innate immune responses. Unlike a plasmid-based system, the RNP system is transient with faster turnover. The result is that it minimizes random integrations and off-target events.
TrueGuide Synthetic gRNA sequences are analyzed by mass spectrometry to verify sequence integrity.
To estimate CRISPR/Cas9 mediated editing efficiency in a pooled cell population, one could use GeneArt Cleavage Detection kit (Part No. A24372), perform next generation sequencing (NGS), or use Sanger sequencing-based analysis. Using NGS of the amplicons from edited populations or Sanger sequencing of amplicons cloned into plasmids gives a more accurate estimate of % editing efficiency and indel types.
TrueGuide Synthetic CRISPR gRNA
Invitrogen TrueGuide Synthetic gRNAs are ready-to-transfect CRISPR guide RNAs designed by a proprietary algorithm for high-efficiency editing. These predesigned, guaranteed guide RNAs offer targeted knockdown with high specificity across the human and mouse genomes. To get started, use the search tool above to search for your gene
Note: TrueGuide crRNA:tracrRNA and unmodified synthetic single guide RNA (sgRNA) have been discontinued. We now only offer our pre-designed modified synthetic sgRNA, which demonstrates superior efficiency across more cell types than the two-part system. Predesigned unmodified sgRNA can still be ordered from the Custom gRNA Order tool. We apologize for any inconvenience this may cause. If you have questions, contact us at techservices@thermofisher.com.
![]() TrueGuide 1-piece modified Synthetic gRNA | |
Predesigned knockouts for mouse and human cells | |
Custom sequences | |
Validated controls available | |
Available in plate format | |
Ready-to-transfect | |
Chemical modifications* | |
Ideal for primary and stem cells |
*Chemical modifications include 2´O-Methyl analogs and phosphorothioate linkages to increase editing efficiency and protect against nuclease degradation.
Lentiviral CRISPR gRNA
Check out our LentiArray CRISPR gRNA if you need to enrich edited cells, are working with difficult-to-transfect cells, or require long-term sgRNA expression.
Cell line-specific transfection conditions using Lipofectamine CRISPRMAX Reagent
The following cell line-specific conditions are provided as a starting point for transfecting wild-type cells with TrueGuide Synthetic gRNA and TrueCut Cas9 Protein v2 using the Lipofectamine CRISPRMAX Transfection Reagent. Further optimization of the transfection conditions may be necessary for best results.
NOTE: Scroll to the right within the table to see all data.
Cell type | Source | Media | Cell seeding density/well (×103) one day before transfection | TrueCut Cas9 Protein v2/gRNA (ng/pmoles) | Lipofectamine Cas9 Plus Reagent/well (μL) | Lipofectamine CRISPRMAX Reagent/well (μL) | ||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Well format | — | — | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well | 96-well | 24-well | 6-well |
HEK293 | Human embryonic kidney | DMEM | 18 | 90 | 450 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.4 | 2 | 10 |
U2OS | Human osteosarcoma | McCoy5A | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
A549 | Human epithelial lung carcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
THP1 | Human peripheral blood monocyte leukemia | RPMI | 10 | 50 | 250 | 400/2.4 | 2000/12 | 10000/60 | 0.8 | 4 | 20 | 0.3 | 1.5 | 7.5 |
K562* | Human leukemia bone marrow | RPMI | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
iPSC* | Human induced pluripotent stem cells | Essential 8 | 8 | 40 | 200 | 300/2 | 1500/10 | 7500/50 | 0.6 | 3 | 15 | 0.3 | 1.5 | 7.5 |
HepG2 | Human hepatocellular carcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
MDA- MB231 | Human epithelial (breast) adenocarcinoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
N2A | Mouse brain neuroblastoma | DMEM | 10 | 50 | 250 | 250/1.5 | 1250/7.5 | 6250/37.5 | 0.5 | 2.5 | 12.5 | 0.3 | 1.5 | 7.5 |
*Use the Neon Transfection System for higher editing efficiency. |
Cell-line specific electroporation conditions using Neon system
The following cell line specific conditions are provided as a starting point for transfecting wild-type cells with TrueGuide Synthetic gRNA and TrueCut Cas9 Protein v2 using the Neon Transfection System 10 μL Kit. Further optimization of the electroporation or nucleofection conditions may be necessary for best results.
Cell type | Source | Media | Number of cells/10-μL reaction (× 103) | TrueCut Cas9 Protein v2/gRNA (ng/pmoles) | Neon electroporation conditions* |
---|---|---|---|---|---|
Well format | — | — | 24-well | ||
HEK293 | Human embryonic kidney | DMEM | 150 | 1250/7.5 | 1150 V/20 ms/2 pulses |
U2OS | Human osteosarcoma | McCoy5A | 150 | 1250/7.5 | 1400 V/15 ms/4 pulses |
A549 | Human epithelial lung carcinoma | DMEM | 120 | 1250/7.5 | 1200 V/20 ms/4 pulses |
THP1 | Human peripheral blood monocyte leukemia | RPMI | 200 | 2000/12 | 1700 V/20 ms/1 pulse (#5) |
K562 | Human leukemia bone marrow | RPMI | 200 | 1250/7.5 | 1700 V/20 ms/1 pulse (#5) |
iPSC | Human induced pluripotent stem cells | Essential 8 | 80 | 1500/10 | 1200 V/20 ms/2 pulses (#14) |
iPSC | Human induced pluripotent stem cells | StemFlex | 80 | 1500/10 | 1200 V/30 ms/1 pulse (#7) |
Human primary T-cell | Healthy donor derived | OpTmizer + 2% human serum | 200 | 1250/7.5 | 1600 V/10 ms/3 pulses (#24) |
Jurkat T-cell | Human peripheral blood lymphocyte | RPMI | 200 | 1250/7.5 | 1700 V/20 ms/1 pulse (#5) |
HepG2 | Human hepatocellular carcinoma | DMEM | 120 | 1250/7.5 | 1300 V/30 ms/1 pulse (#8) |
N2A | Mouse brain neuroblastoma | DMEM | 100 | 1250/7.5 | 1400 V/30 ms/1 pulse (#9) |
*Recommendations for the Neon electroporation settings are based on the culture conditions tested. |
Application data
Up to 90% functional knockout in T-cells
Figure 1. High efficiency functional knockout in T-cells. Human T cells were isolated and activated using Invitrogen Dynabeads magnetic beads (Cat. Nos. 11344D and 11141D, respectively) and then transfected with TrueCut Cas9 Protein v2 and TrueGuide Synthetic gRNA for T cell receptor alpha (TRAC) or beta (TRBC) regions using the Invitrogen Neon delivery system. Following transfection, editing efficiency was measured by (A) the Invitrogen GeneArt Cleavage Detection assay, or by (B,C) functional knockout measured by % T cell receptor negative (TCR–) cells using the Invitrogen Attune NxT Flow Cytometer. Cells analyzed by flow cytometry were stained with T cell receptor–specific antibody conjugated to PE. Here we see up to 90% functional knockout in T cells using our optimized protocol.
Genome editing in cancer cells
Figure 2. Prevail with tools to help unravel the complexities of cancer cells. A wide range of cancer cell lines and gene targets were tested to determine editing efficiencies using the modified single TrueGuide Synthetic gRNA (Cat. No. A35511) complexed with TrueCut Cas9 Protein V2 (Cat. No. A36497). A two-fold increase in efficiency was observed as compared to editing systems from competitors. Here the Cas9 RNP complex was delivered into cell lines indicated using Lipofectamine CRISPRMAX (Cat. No. CMAX00003) and at 72 hours post-transfection, editing efficiency was accessed by Ion Torrent NGS sequencing (NGS) or GeneArt Genomic Cleavage Detection (GCD) Kit (Cat. No. A24372).
Editing efficiencies that outperform the competition
Figure 3. Invitrogen CRISPR genome editing tools offer consistently higher editing efficiency in difficult targets, up to 2 times higher than the competition. Genome editing of multiple gene targets was performed with TrueCut Cas9 Protein V2 and corresponding TrueGuide Synthetic gRNAs. Delivery was achieved using recommended transfection protocols and Lipofectamine CRISPRMAX. The graphs also compare the same experiments using products and recommended protocols another manufacturer. With the Invitrogen system, cleavage efficiency (as measured by %Indel) is improved for low efficient loci (PRKCG-T1 and CMPK1-T1) and shows consistently superior efficiency when compared to competitor products and protocols even for challenging loci.
Cell viability
Figure 4. Optimized protocols designed for minimal cell toxicity. This data set illustrates how the combination of gRNAs synthesized using our proprietary design and delivered using our recommended transfection protocols minimize impact on overall cell viability. Lipofectamine CRISPRMAX were used to complex and deliver TrueCut Cas9 Protein V2 and target specific TrueGuide Synthetic gRNAs for editing several loci including two known to be challenging to edit (PRKCG-T1 and CMPK1-T1). Cell viability was measured by PrestoBlue.
User guide
CRISPR validated protocol collection
Frequently asked questions: TrueGuide Synthetic gRNA
Get your CRISPR guide RNA questions answered
gRNA, or guide RNA, is part of the CRISPR-Cas9 system. The gRNA molecule guides the Cas9 protein to a specific genomic locus via base pairing between the crRNA sequence and the target sequence. The CRISPR-Cas9 system is composed of a short noncoding gRNA that has two components: a target-specific CRISPR RNA (crRNA) and an auxiliary trans-activating crRNA (tracrRNA).
sgRNA stands for single guide RNA. As opposed to the native S. pyogenes CRISPR-Cas9 system that uses a two-part guide (CRISPR RNA [crRNA] hybridized with a tracrRNA), an sgRNA combines both RNAs into a single, long RNA. sgRNA are typically 100 nt in length and can be expressed from a vector or chemically synthesized.
crRNA is one strand of the conventional two-part guide RNA system. The TrueGuide crRNA:tracrRNA two-part guide RNA product format has been discontinued to simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
tracrRNA is one strand of the conventional two-part guide RNA system. The TrueGuide crRNA:tracrRNA two-part guide RNA product format has been discontinued to simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
gRNAs are available as chemically synthesized RNA oligos or generated as in vitro transcribed (IVT) gRNAs using the GeneArt Precision gRNA synthesis kit. The IVT gRNA format can also be purchased as a custom ready-to-use option from GEMServices@thermofisher.com. Chemically synthesized gRNA are available from Thermo Fisher Scientific in three different formats:
- Modified sgRNA format: Invitrogen customers can purchase the single guide RNA (sgRNA), which is comprised of both the crRNA and tracrRNA as a single long RNA with 2′ O-Methyl and phosphorothioate modifications on both 5′ and 3′ ends of the molecule. These modifications enhance editing efficiency by increasing binding to the target site and inhibiting nuclease degradation, respectively.
- Two-part format: The TrueGuide crRNA:tracrRNA two-part guide RNA product format has been discontinued to simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
- Unmodified sgRNA format: Customers can purchase a single guide RNA (sgRNA) without any chemical modifications from the Custom gRNA Order tool. The predesigned option for unmodified sgRNA has been discontinued. To simplify our synthetic gRNA offering and focus on the modified sgRNA, which demonstrates superior efficiency across more cell types than the two-part system.
Chemical modifications include 2’ O-Methyl analogs and phosphorothioate internucleotide linkages at the 5′ and 3′ ends of the molecule. These modifications enhance editing efficiency by increasing binding to the target site and inhibiting nuclease degradation, respectively.
TrueGuide synthetic gRNA along with TrueCut Cas9 Protein V2 offers maximum editing efficiency across broad cell types while minimizing cell toxicity and innate immune responses. Unlike a plasmid-based system, the RNP system is transient with faster turnover. The result is that it minimizes random integrations and off-target events.
TrueGuide Synthetic gRNA sequences are analyzed by mass spectrometry to verify sequence integrity.
To estimate CRISPR/Cas9 mediated editing efficiency in a pooled cell population, one could use GeneArt Cleavage Detection kit (Part No. A24372), perform next generation sequencing (NGS), or use Sanger sequencing-based analysis. Using NGS of the amplicons from edited populations or Sanger sequencing of amplicons cloned into plasmids gives a more accurate estimate of % editing efficiency and indel types.
Can’t find what you are looking for? Contact us at CRISPR@thermofisher.com
For Research Use Only. Not for use in diagnostic procedures.