Evidence map›Paper›PMID 42766154›Full record

ReviewArchives of microbiology2026

Technical limitations of CRISPR-Cas9 genome editing in bacteria: challenges and future directions.

Abdul Qadeer Shabbir, Javeria Idrees, Ali Arif Khan, Iqbal Ahmad Alvi, Saba Sana, Shafiq Ur Rehman, Muhammad Asif

Abstract readReview
PubMed Publisher
In one paragraph

Review in Archives of microbiology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing 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

0 citing papers in PubMed.

No citing paper in PubMed yet.

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

7 authors.

Abdul Qadeer ShabbirDepartment of Microbiology, Dr. Ikram-ul-Haq Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan.
Javeria IdreesDepartment of Microbiology, Dr. Ikram-ul-Haq Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan.
Ali Arif KhanDepartment of Biotechnology, Dr. Ikram-ul-Haq Institute of Industrial Biotechnology, Government College University, Lahore, Pakistan.
Iqbal Ahmad AlviDepartment of Microbiology, Faculty of Medicine, Afghan International Islamic University, Kabul, Afghanistan.
Saba SanaDepartment of Microbiology, Dr. Ikram-ul-Haq Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan.
Shafiq Ur RehmanInstitute of Microbiology and Molecular Genetics, University of the Punjab, Lahore, Pakistan.
Muhammad AsifDepartment of Microbiology, Dr. Ikram-ul-Haq Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan. m.asif@gcu.edu.pk.ORCID http://orcid.org/0000-0002-2542-8842

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The CRISPR-Cas system, which originated as an adaptive immune system in bacteria and archaea, has been repurposed as a precise and programmable tool for genetic manipulation in both prokaryotes and eukaryotes. Its applications in bacteria include targeted genome modifications, antimicrobial resistance studies, functional genomics studies, and the development of engineered strains for industrial and synthetic biology applications. Therefore, the understanding of technical aspects of CRISPR systems and their underlying molecular mechanisms is essential for experimental accuracy, reproducibility, and biosafety. CRISPR editing introduces several challenges in bacteria, including off-target effects, DNA repair limitations, cytotoxicity of Cas nucleases, and host-specific restriction-modification barriers. In addition to these specific challenges, metabolic burden, sgRNA design, and delivery challenges further introduce limitations. Such issues compromise editing efficiency, genomic stability, and cell viability. Recent studies have focused on improved guide RNA design, alternative Cas variants, refined delivery strategies, and host-adapted engineering as promising directions to enhance editing. This review discusses the principal barriers to CRISPR-Cas9 genome editing of bacteria, evaluates the current strategies for addressing these barriers, and highlights emerging approaches aimed at improving the efficiency, reliability and precision in bacterial genome engineering.

Indexed as

BacteriaCRISPR-Cas SystemsGene EditingGenome, BacterialRNA, Guide, CRISPR-Cas SystemsRNA, Guide, CRISPR-Cas SystemsBacterial genome editingCas9 toxicityCRISPR-Cas9Genome editingMultiplex editingPAM specificitysgRNA design

Identifiers

PMID42766154

What OpenQuestion holds

Textmetadata
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 OpenQuestion graph.