Choosing a guide RNA format
When using the CRISPR/Cas9 system, two approaches are possible. You may either use a sgRNA, which combines the variable cr and constant tracr sequences into one long RNA, or you may use a two-part system with separate crRNA and tracrRNA.
One-part RNA (sgRNA)
Single RNA molecule comprised of both crRNA and tracrRNA sequences. Suitable for challenging experimental conditions (e.g., high nuclease environments or with Cas9 mRNA). Contains chemical modifications for the highest level of stability.
Two-part RNAs: CRISPR-Cas9 crRNA
To be used together with a tracrRNA to form a functional guide RNA duplex. Suitable for most applications. Contains chemical modifications to protect from degradation by cellular RNases.
Two-part RNAs: CRISPR-Cas9 crRNA XT
To be used together with a tracrRNA to form a functional guide RNA duplex. Suitable for challenging experimental conditions (e.g., high nuclease environments or with Cas9 mRNA). Contains additional chemical modifications compared to standard crRNA to provide a cost-effective option for increased stability.
IDT's guide RNA formats & vial sizes
| Guide RNA | Size |
|---|---|
| Alt-R® CRISPR-Cas9 sgRNA | 2, 10, 50, 100 nmol |
| Alt-R® CRISPR-Cas9 crRNA | 2, 10, 50, 100 nmol |
| Alt-R® CRISPR-Cas9 crRNA XT | 2 and 10 nmol |
| Alt-R® CRISPR-Cas9 tracrRNA | 5, 20, 100 nmol |
How to design guide RNAs
IDT offers predesigned single guide RNAs and two-part guide RNAs for the following species: human, mouse, rat, zebrafish, nematode. Alternatively, design your custom guide RNA using IDT's design tool or check your existing design using IDT's design checker.
Visit IDT CRISPR-Cas9 design page
Order predesigned guide RNAs Use design tool Check your own design
Choosing a Cas9 enzyme
Cas9 is the the most widely used Cas nuclease. Use Cas9 if this is your first attempt at editing your gene of interest and if your target organism has a standard GC content.
| Cat-No. | Nuclease | Size | Comments |
|---|---|---|---|
| 1081058 | Alt-R® S.p. Cas9 Nuclease V3 | 100 ug | Wild-type version with strong nuclear localization signal (NLS). High on-target efficiency due to effective delivery to the nucleus. |
| 1081060 | Alt-R® S.p. HiFi Cas9 Nuclease V3 | 100 ug | Improved version with increased specificity. High on-target efficiency paired with strongly reduced off-target activity. Use when off-target modifications are of concern. |
| 1081062 | Alt-R® S.p. Cas9 D10A Nickase V3 | 100 ug | This Cas9 D10A Nickase V3 creates a single-stranded cut in the targeted strand of DNA. |
| 1081064 | Alt-R® S.p. Cas9 H840A Nickase V3 | 100 ug | This Cas9 H840A Nickase V3 creates a single-stranded cut in the non-targeted strand of DNA. |
| 1081066 | Alt-R® S.p. dCas9 Protein V3 | 100 ug | Enzymatically “dead” Cas9 mutant, lacks cleavage activity on both DNA strands. |
CRISPR-Cas9 Control Kits
Separate kits for human, mouse, or rat. Includes tracrRNA, HPRT positive control crRNA, negative control crRNA#1, HPRT Primer Mix, and Nuclease-Free Duplex Buffer. Purchase the Cas9 enzyme (or expression plasmid) and PCR reagents separately.
| Cat-No. | Kit | Size | Comments |
|---|---|---|---|
| 1072554 | Alt-R® CRISPR-Cas9 Control Kit, human | 2 nmol | Contains control sequences specific to human. |
| 1072555 | Alt-R® CRISPR-Cas9 Control Kit, mouse | 2 nmol | Contains control sequences specific to mouse. |
| 1072556 | Alt-R® CRISPR-Cas9 Control Kit, rat | 2 nmol | Contains control sequences specific to rat. |
Friendly CRISPR Clash Webinar
First Back2Back Academic and Industrial CRISPR Webinar in Switzerland.
- Dissecting the molecular mechanism of Cas9 specificity
Speaker: Dr. Martin Pačesa, Group of Prof. Bruno Correia, EPFL - Laboratory of Protein Design and Immunoengineering
- Improved methods for CRISPR HDRresearch using Alt-R™ modified donors and Alt-R HDR Enhancer V2
Speaker: Jennifer Stott, Commercial and Technical Specialist CRISPR & Functional Genomics - IDT
For research use only. Not for use in diagnostic procedures. Unless otherwise agreed to in writing, IDT does not intend for these products to be used in clinical applications and does not warrant their fitness or suitability for any clinical diagnostic use. Purchaser is solely responsible for all decisions regarding the use of these products and any associated regulatory or legal obligations.