The study identifies technical challenges associated

The gap

Evidence profile

Sourced from the future work and stated challenges of the source papers, classified as general, spanning 3 journals.

Supporting evidence — 3 representative gaps

• CRISPR/Cas9 in perspective: evaluating efficacy, delivery methods, and ethical challenges in genome editing (2026) · Molecular Biology Reports · doi

  CRISPR/Cas9 technology has significantly transformed the fields of genetics and genomics by providing a versatile and programmable platform for genome engineering. Its ability to introduce targeted genetic modifications has enabled the development of improved disease models, genetically modified organisms with desirable traits, and emerging therapeutic strategies for a variety of genetic disorders. Consequently, CRISPR-based genome editing has become an important tool in both fundamental biological research and translational biomedical applications. Despite these advances, several challenges remain that limit the widespread clinical implementation of CRISPR technologies. Ensuring the safety and precision of genome editing is essential, particularly in therapeutic contexts where unintended genomic alterations may have long-term biological consequences. Off-target mutations, insertion–deletion events (INDELs), and frameshift mutations continue to represent important technical challenges that require careful monitoring and methodological refinement. Recent developments in RNA-targeting CRISPR systems also highlight the expanding capabilities of genome engineering technologies. For example, programmable RNA acetylation systems based on CRISPR–Cas13, such as the engineered eNAT10– dCas13 fusion protein, have been developed to modulate RNA modifications, including N4-acetylcytidine (ac4C), 1 3Molecular Biology Reports (2026) 53:719  719 Page 14 of 20 s e c n e r e f e R ] 3 1 1, 1 1 1 [ d e t r o p e R s n o i t a t u m e v i t a n r e t l a () m e t s y s t e g r a t - ff O s n o i t a t u m e n e g r o, y c n e i c ffi e R D H, y c n e u q e r f l e d n i,. g. e ( s c i r t e m e m o c t u o, s e i t i l a d o m y r e v i l e d, s t e g r a t e n e g g n i d u l c n i, s n o i t a c i l p p a l a t n e m / i r e p x e 9 s a C R P S I R C d e t r o p e r f o y r a m m u s e v i s n e h e r p m o C 1 e l b a T s m e t s y s g n i s n o i t a t u m - t i d e e v i t a n r e t l A s n o i t a t i m i l l a t n e m i r e p x E d e v r e s b O d o h t e m y r e v i l e D t e g r a T s u c o l l e d o m e n e g t e g r a T l a t n e m i r e p x E e v i t c e j b O o N S.

  generalfuture workevidence 5/5
  Keywords: crispr genome programmable engineering genetic modifications therapeutic based editing important challenges technolo gies mutations systems
• CRISPR-screen informed engineered T cell therapies (2026) · Frontiers in Immunology · doi

  CRISPR screens have mapped the core circuitry by which T cells achieve anti-tumor activity, repeatedly converging on JAK/STAT cytokine signaling, proximal and distal TCR/NF-kB pathways including RNA stability programs, and transcriptional/epigenetic checkpoints that shape differentiation and persistence. Looking forward, a stage-aligned, context-aware-design strategy as presented herein provides a practical approach for the selection of targets and pathways to optimally enhance T cell therapies. As dual editing of identified target combinations can integrate complementary bene- fits into T cell therapies, combinatorial editing will be an important tool in this approach. These insights provide a rational foundation for engineering adoptive cell therapies.

  generalfuture workevidence 5/5
  Keywords: approach cell therapies crispr tumor pathways editing screens mapped core circuitry cells achieve anti activity
• CRISPR/Cas9 knockout of N-acetyltransferase 1 influences mesenchymal transition in glioblastoma in a cell line dependent manner (2026) · Journal of Cancer Research and Clinical Oncology · doi

  The study identifies technical challenges associated with CRISPR/Cas9 knockout, including off-target effects and variable knockout efficiency. The research highlights domain challenges, such as the complexity of glioblastoma and the need for personalized treatment approaches. The study notes the challenge of interpreting the results of the CRISPR/Cas9 knockout experiments in the context of glioblastoma.

  generalstated challengesevidence 5/5
  Keywords: study identifies technical challenges associated crispr cas9 knockout

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