Evidence map›Paper›PMID 40966510›Full record

ArticleNucleic acids research2025

AsCas12a tolerates insertions in target DNA.

Santosh R Rananaware, Grace M Shoemaker, Brianna L M Pizzano, Emma K Vesco, Luke Samuel W Sandoval, Jordan G Lewis, August P Bodin, Sarah J Flannery, Ian H Lange, Dheeraj Pedada and 8 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Nucleic acids research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

18 authors.

Santosh R RananawareDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Grace M ShoemakerDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Brianna L M PizzanoDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Emma K VescoDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Luke Samuel W SandovalDepartment of Computer Science, University of Florida, Gainesville, FL 32611, United States.
Jordan G LewisDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
August P BodinDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Sarah J FlanneryDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Ian H LangeDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Dheeraj PedadaDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Anne FangDepartment of Chemical Biology, University of Florida, Gainesville, FL 32610, United States.
Sydney G AntalDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Daisy AguilarDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Noah R RakestrawDepartment of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610, United States.
Vedant N KaralkarDepartment of Computer Science, University of Florida, Gainesville, FL 32611, United States.
Katelyn S MeisterDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Long T NguyenDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.
Piyush K JainDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611, United States.ORCID 0000-0001-7153-442X

Funding

Discovery and engineering of CRISPR/Cas systemsR35GM147788 · NIGMS · UNIVERSITY OF FLORIDA · PI Piyush K Jain · 2022 to 2026
$1.8M
A rapid CRISPR-based self-testing platform for early detection of HIVR61AI181016 · NIAID · UNIVERSITY OF FLORIDA · PI Piyush K Jain · 2024 to 2026
$1.3M
Rapid detection of Hepatitis C virus using CRISPR/CasR21AI156321 · NIAID · UNIVERSITY OF FLORIDA · PI JAIN, PIYUSH K · 2021 to 2022
$419k
Rapid point-of-care detection of Hepatitis C viral RNA using multiplexed CRISPR/Cas platformsR21AI168795 · NIAID · UNIVERSITY OF FLORIDA · PI JAIN, PIYUSH K · 2022 to 2023
$419k
Exxon Mobil Gator Alumni FacultyNIAID NIH HHS 5R61AI181016NIAID NIH HHS R21 AI156321NIAID NIH HHS R21 AI168795NIAID NIH HHS R21AI168795NIAID NIH HHS R35GM147788NIAID NIH HHS R61 AI181016NIGMS NIH HHS R35 GM147788NIH HHS R21AI156321Shah FoundationUF Herbert Wertheim College of EngineeringUniversity of Florida
6 · The paper itself

Abstract

CRISPR-Cas12a enzymes are RNA-guided nucleases widely used for programmable genome editing and diagnostics. Perfect complementarity between guide RNA and target DNA is essential for efficient binding and cleavage by Cas12a. However, we report that a particular ortholog of Cas12a, Acidaminococcus sp. Cas12a (AsCas12a), shows an unexpected tolerance to noncomplementary insertions at various positions in its DNA target. AsCas12a remains functional despite DNA bubbles or loops in the CRISPR-RNA (crRNA)-target DNA duplex, displaying both cis- and trans-cleavage activities even when the target harbors insertions of lengths 1-20 nucleotides in the crRNA-binding region. This activity is sequence-independent and works for ssDNA and is observed on dsDNA in vitro for specific insertion lengths/positions and DNA topologies but is strongly diminished in cells. Among 12 Cas12a orthologs tested, only AsCas12a exhibits this tolerance, making it a unique member of the Cas12a family. Structural analysis suggests a distinctive α-helix in AsCas12a's WED domain is required for this flexibility. Upon deleting this α-helix, AsCas12a loses its ability to tolerate insertions. This discovery can be utilized to detect single-nucleotide polymorphisms and enable protospacer adjacent motif (PAM)-flexible DNA cleavage with Cas12a. Our findings expand our fundamental understanding of CRISPR-Cas12a systems. In conclusion, we uncover and characterize a unique property of AsCas12a to tolerate insertions in its target.

Indexed as

AcidaminococcusBacterial ProteinsCRISPR-Associated ProteinsCRISPR-Cas SystemsDNAEndodeoxyribonucleasesDNA CleavageGene EditingModels, MolecularRNA, Guide, CRISPR-Cas SystemsBacterial ProteinsCas12a proteinCRISPR-Associated ProteinsDNAEndodeoxyribonucleasesRNA, Guide, CRISPR-Cas Systems

Identifiers

PMID40966510
PMCPMC12448901

What Socratic holds

Textmetadata
LicenceCC BY-NC
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.