Custom CRISPR gRNA ordering

The Alt-R CRISPR-Cas9 gRNA products have been updated!

The Alt-R CRISPR gRNA products (crRNA:tracrRNA and sgRNA) are now chemically modified for enhanced RNA stability. To read more about the latest performance improvements, visit the product page.

The Alt-R CRISPR-Cas9 system now offers additional guide RNA options:

• Alt-R crRNA: the most popular and economical option for CRISPR experiments (must be used with the universal Alt-R tracrRNA)
• Alt-R crRNA XT: enhanced modifications to provide additional stability and high performance (must be used with the universal Alt-R tracrRNA)
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• Alt-R sgRNA: chemically modified single guide RNAs can provide the best results for the most challenging cell types and applications

Simplified ordering

Enter the 20 base DNA sequence upstream of the PAM site, as shown, and you are ready to continue. The ordering tool will automatically convert the DNA sequence to RNA during the ordering process. If you are pasting your CRISPR target site from an online design tool, make sure you verify the correct strand orientation.

Instructions for ordering Cas12a (Cpf1) crRNAs

Enter the 20–24 base DNA sequence 3′ of the target Cas12a (Cpf1) PAM site (TTTN), and you are ready to continue. The ordering tool will automatically convert the DNA sequence to RNA during the ordering process. If you are pasting your Cas12a (Cpf1) target site from an online design tool, make sure you verify the correct strand orientation.

Enter your DNA sequence in the Sequence box and use the drop down menus to select your desired specifications. We provide three options for Alt-R HDR Donor Oligo modifications. Alt-R HDR modified is our recommended option for the best HDR performance and includes 2 phosphorothioate (PS) linkages and also an IDT proprietary end-blocking modification at each end to provide increased stability. Phosphorothioate modified will contain 2 PS linkages at each end of the DNA sequence and Unmodified will contain only standard DNA.

For optimal synthesis quality, guide RNA sequences within the same plate should share similar lengths and modifications.

• Generate custom novel CRISPR guide RNAs (such as pegRNA, and Cas13a gRNA) targeting any sequence from any species.
• Generate guide RNAs of any length from 30 – 150 nt, with the below base and modification options.
• Available base types and modifications are:
  • RNA, expressed as 'r_', for example rA, rU
  • 2' O-methyl RNA bases are entered as 'm_'
  • Phosphorothioated RNA bases are entered as 'r_*'
  • Phosphorothioated 2'-O-methyl RNA bases are entered as 'm_*'
  • 5’ and 3’ Alt-R end-blocking modification entered as /AltR1/ and /AltR2/
• Sequence will be synthesized exactly as entered.

Example sequences:

1. rCrGrUrUrArArUrCrGrCrGrUrArUrArArUrArCrGrGrUrUrUrUrArGrArGrCrUrArUrGrCrU
2. /AltR1/rCrGrUrUrArArUrCrGrCrGrUrArUrArArUrArCrGrGrUrUrUrUrArGrArGrCrUrArUrGrCrU/AltR2/
3. mA*mG*mC*rArUrArGrCrArArGrUrUrArArArArUrArArGrGrCrUrArGrUrCrCrGrUrUrArUrCrArArCrUrUrGrArArArArArGrUrGrGrCrArCrCrGrArGrUrCrGrGrUrGrC*mU*mU*mU

• Order 2'MOE or Affinity Plus ASOs of any length from 14 - 24 nt, with the below base and modification options.
• Available ASO 2'MOE base types and modifications are:
  • Phosphorothioated DNA bases are entered as '_*'
  • 5' 2′-MOE modified DNA bases entered as /52MOEr_/
  • Internal 2′-MOE modified DNA bases entered as /i2MOEr_/
  • 3' 2′-MOE modified DNA bases entered as /32MOEr_/
• Available ASO Affinity Plus base types and modifications are:
  • Phosphorothioated DNA bases are entered as '_*'
  • Affinity Plus entered as '+_'
• Sequence will be synthesized exactly as entered.

Example sequences:

1. /52MOErT/*/i2MOErC/*/i2MOErC/*/i2MOErT/*/i2MOErG/*C*G*A*A*A*T*G*T*C*C*/i2MOErA/*/i2MOErT/*/i2MOErC/*/i2MOErG/*/32MOErT/
2. /52MOErT/*/i2MOErC/*/i2MOErC/*/i2MOErT/*C*G*A*A*A*T*G*T*C*C*/i2MOErT/*/i2MOErC/*/i2MOErG/*/32MOErT/
3. +C*+T*+G*C*G*A*A*A*T*G*T*C*C*+A*+T*+C
4. +C*+T*C*G*A*A*A*T*G*T*C*+A*+T*+C

• Generate custom Antisense oligonucleotides (ASOs) of any length from 12 - 36 nt, with the below base and modification options.
• Available base types and modifications are:
  • 2' O-methyl RNA bases are entered as 'm_'
  • Phosphorothioated DNA bases are entered as '_*'
  • Affinity Plus entered as '+_'
  • Phosphorothioated 2'-O-methyl RNA bases are entered as 'm_*'
  • 5’ Methyl dC modification entered as /5Me-dC/
  • 5’, Internal, and 3' 2′-MOE modified DNA bases entered as /52MOEr_/, /i2MOEr_/, and /32MOEr_/
• Sequence will be synthesized exactly as entered.

Example sequences:

1. T*C*C*T*G*C*G*A*A*A*T*G*T*C*C*A*T*C*G*T
2. /52MOErT/*/i2MOErC/*/i2MOErC/*/i2MOErT/*/i2MOErG/*C*G*A*A*A*T*G*T*C*C* /i2MOErA/*/i2MOErT/*/i2MOErC/*/i2MOErG/*/32MOErT/
3. +C*+T*+G*C*G*A*A*A*T*G*T*C*C*+A*+T*+C
4. mU*mC*mC*mU*mG*C*G*A*A*A*T*G*T*C*C*mA*mU*mC*mG*mU