Enhance your Gene Target efficiency using Synthetic Long RNAs for CRISPR/Cas9

CRISPR (clustered regularly interspaced short palindromic repeat) loci and a diverse cassette of CRISPR-associated (cas) genes are part of an adaptive immune system found in bacteria and archaea. The CRISPR/Cas9 system based on bacterial CRISPR-associated protein-9 nuclease (Cas9) from Streptococcus pyogenes is a new gene editing tool used in cell and molecular biology. Long RNA targeting efficiency is an important aspect in genome editing. Bio-Synthesis provides chemically synthesized long RNA and RNA transcripts that can be used to improve target specificity.

Bio-Synthesis has been successful using chemical and enzymatic synthesis for long RNAs of up to 400 bases. RNA oligos between 80-400 bases are synthesized using the latest nucleic acid technology and purified using RNase free PAGE electrophoresis combined with HPLC. Our RNA synthesis services include mass spectrometry analysis. Purified RNA oligos undergo stringent analytical HPLC or gel electrophoresis to determine final purity.

The Enzymatic method (IVT) allows very long sequences (3,000 bases or higher) to be generated at high yields. Bio Synthesis has successfully synthesized thousands of semi-synthetic long RNA transcripts using proprietary methods to optimize RNA manufacturing in vitro using SP6 or T7 polymerase. Each in vitro transcribed long RNA oligo is meticulously monitored during synthesis and controlled according to Bio Synthesis’s stringent quality assurance and quality control standards.

Chemically Synthesized Long RNA include:

• Unmodified & Modified RNA: Deprotected, desalted and ready to use, Fast delivery and high throughput capacity
• Modified bases: 5’ Amino Modifier C12, 2’O-Methyl RNA bases, 5' 5-Methyl dC, Int 5-Methyl dC, 3’Phosphorylation
• Process optimization: For scales up to multi-gram quantity
• High Quality: RNase free PAGE purified and packaged in an inert chemical matrix

Long RNA Synthesized by In Vitro Transcription Include:

• Modified bases: 5-Me-CTP, pseudo-UTP and other modified NTPs
• Terminal Capping: m7G(5')ppp(5')G (mCAP, Anti Reverse Cap Analog (ARCA), poly A or poly(dT), fluorescent dye incorporation.
• Other modifications: Poly A or poly(dT), fluorescent dye incorporation.
• High Quality: Column based purification, HPLC or PAGE delievered in a RNaseSAFE^TM Tube
• Flexible Solutions: Transcripts from plasmids, PCR products, oligonucleotides and cDNA.

Please note that creation and use of synthetic oligos for CRISPR applications requires a license from ERS genomics, the holders of IP for CRISPR technology. We do not have a license to provide synthetic oligos used in CRISPR technology from ERSGenomics. Contact ERSGenomics.com for further information.