Beyond CRISPR: searchRNA provides a new method for precise gene editing

Scientists at the University of Sydney have developed a more precise and flexible genetic analysis tool than the industry standard, CRISPR, which is revolutionizing engineering in medicine, agriculture and biotechnology.

SeekRNA uses an integrable strand of ribonucleic acid (RNA) that can precisely identify sites for insertion into the sequence, facilitating the control and reduce errors.

The new genetic testing tool is being developed by a team led by Dr. Sandro Ataide in the School of Life and Environment. Their findings are published in Nature Communication.

“We are very excited about the potential for this technology. SeekRNA’s ability to target selectivity with precision and flexibility sets the stage for a new era of genetic engineering, surpassing the limitations of technology. now,” said Dr. Ataide.

“With CRISPR you need additional components to have a ‘cut-and-paste tool,’ but the promise of seekRNA is an independent ‘cut-and-paste tool’ with high accuracy possible then give more parts. DNA sequence.”

CRISPR relies on creating a break in both strands of the target DNA, the double-helix genetic code of life, and requires other proteins or the replication machinery. DNA modification to introduce new DNA sequence. This can cause errors.

Said Dr. Ataide, “SearchRNA can directly attach the base and attach the new DNA without the use of any other protein. This allows for cleaner control tools and more accurate and few mistakes.”

Gene-editing has opened up completely new areas of research and use since the development of CRISPR more than 10 years ago. It has led to the improvement of disease control in fruits and vegetables, reducing the cost and speed of diagnosing human diseases, helping in the search for a cure for sickle cell disease and allowing for the development of cancer treatments. new called (CAR) T- cell therapy.

“We are very much in the early stages of what gene editing can do. We hope that by developing this new approach to gene editing, we can contribute to the advancement of life. health, agriculture and biotechnology,” said co-author Professor Ruth Hall from the University of Sydney.

Correct generation calculations

SeekRNA is derived from a family of naturally occurring transcription factors called IS1111 and IS110, found in bacteria and archaea (cells that do not have a nucleus). Most sequence binding proteins show little or no target selectivity, but these families show high specificity.

This is exactly what searchRNA has used to achieve its desired results so far. Using precision from the insertion sequence of the family, find RNA that can replace any genomic sequence and insert the new DNA in a correct order. zero.

“In the laboratory we have successfully studied searchRNA in bacteria. Our next step is to study whether the technology can be adapted for the more complex eukaryotic cells found in humans,” said Dr. Ataide.

An advantage of the method reported in this study is that it can be used with only a single protein of small size with a hole searchRNA strand. In short, it activates the genetic material. SeekRNA is made up of a small protein of 350 amino acids and an RNA strand of between 70 and 100 nucleotides.

A similar system can be incorporated into biological vehicles (vesicles or lipid nanoparticles) for delivery to cells of interest.

Direct insertion into DNA

Another point of difference is the ability of this technology to insert DNA sequences into the desired location on its own, something that cannot be done with many instruments. maybe.

“Current CRISPR technologies have limitations on the number of gene sequences that can be transferred,” said University of Sydney researcher Rezwan Siddiquee, the author of the paper. “This limits the scope of the application.”

Around the world, other teams are pursuing similar studies using IS1111 and IS110 family genes. However, said Dr. Ataide they showed results for one member of the IS110 family and relied on a much larger RNA. The team in Sydney is expanding its methods through direct laboratory experiments and using searchRNA itself.