Back to methods browser

First-pass extracted concept

multiplex CRISPR/Cas9 genome editing

Candidate: toolkit itemType: engineering method1 source documents2 linked claims
Jump to explainersJump to evidenceJump to sourcesJump to claims
Live refresh every 5sNext refresh in 5s

Aliases

CRISPR/Cas9, multiplex CRISPR/Cas9

Extracted Explainers

What the tool is doing

This method was used to simultaneously edit five α-1,3-fucosyltransferase genes and two β-1,2-xylosyltransferase genes in Nicotiana benthamiana. The stated aim was to generate glycoengineered plants with more humanized glycosylation profiles.

Source 1DOIPubMed

Resources required

The abstract supports the need for CRISPR/Cas9 editing, transformed plant lines, genotyping of T0 transformants, and screening of T1 and T2 generations for Cas9-free homozygous plants.

Source 1DOIPubMed

What problem it solves

It addresses plant-specific α-1,3-fucosylation and β-1,2-xylosylation, which the paper describes as immunogenic and as barriers to plant-based therapeutic protein production.

Source 1DOIPubMed

What it does not solve

The abstract does not show that editing alone produces a final therapeutic product or resolves all downstream manufacturing and regulatory issues beyond the targeted glycosylation problem.

Source 1DOIPubMed

Alternatives

The web research summary identifies TALENs and RNAi as earlier alternative approaches used in related plant glycoengineering work.

Source 1DOIPubMed

Evidence Snippets

We applied multiplex CRISPR/Cas9 genome editing in Nicotiana benthamiana to simultaneously target five α-1,3-fucosyltransferase genes and two β-1,2-xylosyltransferase genes.
Evidence 1Source 1DOIPubMedprovenance

Supporting Sources

Linked Claims

Claim 1editing outcomesupports2025Source 1DOIPubMed

Two T0 lines, HL40 and HL64, showed successful edits in all seven target genes.

Quoted textsource-backed
Two T0 lines (HL40 and HL64) exhibited successful edits in all seven target genes, with mutations consisting of single-base insertions and deletions up to 26 bp.
Claim 2method applicationsupports2025Source 1DOIPubMed

Multiplex CRISPR/Cas9 was applied in Nicotiana benthamiana to simultaneously target five α-1,3-fucosyltransferase genes and two β-1,2-xylosyltransferase genes.

Quoted textsource-backed
We applied multiplex CRISPR/Cas9 genome editing in Nicotiana benthamiana to simultaneously target five α-1,3-fucosyltransferase genes and two β-1,2-xylosyltransferase genes.