Scientists at Kolkata’s Bose Institute have engineered GlowCas9, a bioluminescent CRISPR protein that enables real-time visualisation of genome editing inside living cells.

About Gene Therapy and CRISPR-Cas9

• Gene Therapy Potential: Gene therapy can be a permanent cure for many life-threatening hereditary diseases, but developing effective, affordable, and safe methods remained a challenge for decades.
• CRISPR-Cas9 Mechanism: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) uses a guide RNA to direct the Cas9 enzyme to a specific DNA sequence, where it precisely cuts the DNA to correct genetic defects.
• Limitation in Observation: Scientists could not observe Cas9 functioning in living cells in real time, and 
  • Existing detection methods required either fixing or breaking open cells, making it impossible to track the process as it unfolded.
• Need for Real-Time Monitoring: Tracking gene editing as it happens can help monitor CRISPR operations in living cells and tissues without destroying them.

About GlowCas9

• GlowCas9 is a CRISPR protein that lights up while performing gene editing inside living systems.
• Design: GlowCas9 was designed by Arkadeep Karmakar by fusing Cas9 with a split nano-luciferase enzyme derived from deep-sea shrimp proteins.
• Light Emission Process: The inactive enzyme pieces reconnect when Cas9 folds correctly, producing light.
• This is because when the pieces are brought to proximity, they can reassemble to restore enzymatic activity and produce a visible signal akin to the gentle light of fireflies.
• Real-Time Visualisation Capability: This glowing activity allows monitoring of CRISPR operations in living cells, tissues, and plant leaves without causing harm.

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Advantages of GlowCas9

• Thermostability: GlowCas9 is highly stable and maintains its structure and activity at higher temperatures than the conventional enzyme.
  • Such stability is essential for gene therapy, where stable Cas9 delivery increases treatment success.
• Enhanced DNA Repair Accuracy: GlowCas9 increases the precision of homology-directed repair (HDR), a DNA repair process crucial for fixing hereditary mutations linked to genetic diseases such as sickle cell anaemia, muscular dystrophy, and more.
• Tracking in Plants: GlowCas9 can also be tracked in plant systems. This hints at potential safe, non-transgenic applications in crop improvement.
• Theratracking: GlowCas9 represents a leap in theratracking—the ability to visualise molecular gene therapy in motion.
  • It brings us closer to a future where scientists not only edit genes but also watch the cure unfold, one photon at a time.

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