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 s

The gap

Consensus across the literature

Supporting evidence — 3 representative gaps

• Advancing Functional Genomics in Marchantia polymorpha: A Review of Induced Mutagenesis Toolkits and Applications (2026) · doi

  The diverse mutagenesis tools available for M. polymorpha each have distinct strengths and ideal applications, summa- rized in Table 1. The choice of method depends on the spe- cific research goal. For unbiased phenotype-driven screens, random mutagenesis approaches are indispensable. Heavy- ion beam irradiation currently offers the highest efficiency and diverse mutation spectrum. For testing hypotheses about specific genes, targeted editing tools like CRISPR/Cas9 are unrivalled for their speed and precision in generating knock- outs. While still developing, base and prime editing repre- sent the future for introducing specific nucleotide changes to study protein function and for trait engineering. Abbreviations: EMS, ethyl methanesulfonate; NaN ₃, sodium azide; MNU, N-methyl-N-nitrosourea; LET, linear energy transfer; DSBs, double-strand breaks; gRNA, guide RNA; NHEJ, non-homologous end joining; HDR, homol- ogy-directed repair; CRISPR, clustered regularly interspaced short palindromic repeats; TALENs, transcription activator- like effector nucleases; T-DNA, transferred DNA Several exciting frontiers lie ahead. First, the community must develop standardized, optimized protocols for random mutagenesis (e.g., EMS, heavy-ion) to build large, publicly available mutant libraries. Second, efforts to enhance HDR efficiency and adapt base/prime editing for Marchantia will unlock precise genome engineering. Third, addressing transgene expression variability through synthetic biology approaches will be crucial for complex genetic manipula- tions. Finally, integrating these mutagenesis tools with high- throughput phenotyping and sequencing technologies will enable systems-level approaches to understand gene function in this evolutionarily pivotal plant.

  Keywords: mutagenesis tools approaches editing diverse available random heavy efficiency specific like crispr base prime function
• CRISPR/Cas9 in perspective: evaluating efficacy, delivery methods, and ethical challenges in genome editing (2026) · doi

  CRISPR/Cas9 technology has significantly transformed the fields of genetics and genomics by providing a versatile and programmable platform for genome engineering. Its abil- ity 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 [148]. 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 technolo- gies. Ensuring the safety and precision of genome editing is essential, particularly in therapeutic contexts where unin- tended genomic alterations may have long-term biologi- cal 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 devel- opments in RNA-targeting CRISPR systems also highlight the expanding capabilities of genome engineering technolo- gies. 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 . s e g n e l l a h c l a c i n h c e t d e t a i c o s s a d n a , ) y c n e i c ffi e n o i s s e r p e r s n o i t a t u m s r o t i d e s e s n o p s e r e n u m m i s n o i t a t u m g n i n i a t n o c s e l c i t ] 3 1 1 [ l e d n I e s a b e n i n e d A l l e c - T f o n o i t a v i t c A t e g r a t - ff O - r a p o n a n d i p i L 8 n o x E 9 K S C P s e u q a c a m s e t a m i r p n a m u h - n o n s u s e h R n i g n i t i d e e m o n e G - d a e l s l l e c d e t i d e t s n i a g a d n a n o i t a m m a fl n i o t g n i y c n e i c ffi e g n i t i

  Keywords: crispr genome programmable engineering genetic modifications therapeutic based editing important challenges technolo gies mutations systems
• CRISPR-screen informed engineered T cell therapies (2026) · 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. Coupling CRISPR screen-informed enhancements with the personalized and tumor-reactive features of TIL serves as a prom- ising approach to overcome the current efficacy challenges

  Keywords: approach cell therapies crispr tumor pathways editing screens mapped core circuitry cells achieve anti activity