Nature Plants, Год журнала: 2025, Номер unknown
Опубликована: Апрель 1, 2025
Язык: Английский
Plant Biotechnology Journal, Год журнала: 2025, Номер unknown
Опубликована: Янв. 12, 2025
Summary The Cas12j‐8 nuclease, derived from the type V CRISPR system, is approximately half size of Cas9 and recognizes a 5′‐TTN‐3′ protospacer adjacent motif sequence, thus potentially having broad application in genome editing for crop improvement. However, its efficiency remains low plants. In this study, we rationally engineered both crRNA nuclease. markedly improved When combined, they exhibited robust activity soybean rice, enabling target sites that were previously uneditable. Notably, certain sequences, was comparable to SpCas9 when targeting identical it outperformed Cas12j‐2 variant, nCas12j‐2, across all tested targets. Additionally, developed cytosine base editors based on Cas12j‐8, demonstrating an average increase 5.36‐ 6.85‐fold base‐editing (C T) compared with unengineered system plants, no insertions or deletions (indels) observed. Collectively, these findings indicate hypercompact CRISPR/Cas12j‐8 serves as efficient tool mediated by nuclease cleavage
Язык: Английский
Процитировано
0
bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown
Опубликована: Фев. 12, 2025
TnpB is an evolutionarily diverse family of RNA-guided endonucleases associated with prokaryotic transposons. Due to their small size and putative evolutionary relationship Cas12s, holds significant potential for genome editing mechanistic exploration. However, most TnpBs lack robust gene-editing activity, unbiased profiling mutational effects on activity has not been experimentally explored. Here, we mapped comprehensive sequence-function landscapes a ribonucleoprotein discovered many activating mutations in both the protein RNA. Single-position changes RNA outperform existing variants, highlighting utility systematic scaffold mutagenesis. Leveraging landscape protein, identified enhanced variants from combinatorial library mutations. These increased human cells N. benthamiana by over two-fold fifty-fold relative wild-type TnpB, respectively. In total, this study highlights unknown elements critical regulation endonuclease RNA, reveals surprising amount latent accessible through mutation.
Язык: Английский
Процитировано
0
New Phytologist, Год журнала: 2025, Номер unknown
Опубликована: Март 8, 2025
Genome editing holds great promise for enhancing crop traits; however, progress has been slow due to inefficient delivery methods and reliance on tissue culture regenerating edited plants, which are time-consuming labor-intensive. To address these limitations, innovative culture-free techniques have developed, including meristem through biolistic-mediated RNA virus-mediated delivery. New de novo gene-edited induction root suckering-based cut-dip-budding also established. While approaches show promise, each faces challenges that must be addressed practical application in improvement. We discuss the transformative potential of improvement emphasize need ongoing research refine them maximize their agricultural impact.
Язык: Английский
Процитировано
0
Plants, Год журнала: 2025, Номер 14(6), С. 889 - 889
Опубликована: Март 12, 2025
Recent years have witnessed remarkable progress in plant biology, driven largely by the rapid evolution of CRISPR/Cas-based genome editing (GE) technologies. These tools, including versatile CRISPR/Cas systems and their derivatives, such as base editors prime editors, significantly enhanced universality, efficiency, convenience functional genomics, genetics, molecular breeding. However, traditional genetic transformation methods are essential for obtaining GE plants. depend on tissue culture procedures, which time-consuming, labor-intensive, genotype-dependent, challenging to regenerate. Here, we systematically outline current advancements simplifying GE, focusing optimization process through developmental regulators, development planta methods, establishment nanomaterial- viral vector-based delivery platforms. We also discuss critical challenges future directions achieving genotype-independent, culture-free GE.
Язык: Английский
Процитировано
0
Nature Plants, Год журнала: 2025, Номер unknown
Опубликована: Апрель 1, 2025
Язык: Английский
Процитировано
0