Source 1primary paper2022
Toolkit/split pegRNA prime editors
split pegRNA prime editors
Also known as: SnPEs, split pegRNA prime editors
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Split pegRNA prime editors (SnPEs) are prime editing constructs in which a modified pegRNA is divided into an sgRNA and a separate prime RNA. In the cited 2022 study, this split architecture maintained prime editing activity while increasing design flexibility.
Usefulness & Problems
Why this is useful
SnPEs are useful for reconfiguring pegRNA architecture when the 3-prime extension of a conventional pegRNA may impair RNA stability or folding. The reported split design increases flexibility while preserving prime editing activity in the cited study.
Source:
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Problem solved
These constructs address the design problem that the 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and thereby compromise prime editing activity. Splitting the modified pegRNA into an sgRNA and a prime RNA provides an alternative architecture intended to mitigate that constraint.
Problem links
Need conditional recombination or state switching
DerivedSplit pegRNA prime editors (SnPEs) are prime editing constructs in which a modified pegRNA is divided into an sgRNA and a separate prime RNA. In the cited 2022 study, this split architecture maintained prime editing activity while increasing design flexibility.
Need controllable genome or transcript editing
DerivedSplit pegRNA prime editors (SnPEs) are prime editing constructs in which a modified pegRNA is divided into an sgRNA and a separate prime RNA. In the cited 2022 study, this split architecture maintained prime editing activity while increasing design flexibility.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Techniques
No technique tags yet.
Target processes
editingrecombinationImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulatorswitch architecture: split
SnPEs were created by splitting modified pegRNAs into two RNA components: an sgRNA and a prime RNA. The same study also describes stem-loop aptamer addition at the 3-prime end of pegRNA and tethering strategies to Cas9 nickase, but the supplied evidence does not specify the exact construct requirements needed for all SnPE implementations.
The provided evidence does not report quantitative editing efficiencies, target scope, cell types, or comparative performance across loci. Independent replication and broad validation are not established from the supplied material.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
Stem-loop PEs can be tethered to Cas9 nickase to produce tethered PEs.
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Stem-loop PEs can be tethered to Cas9 nickase to produce tethered PEs.
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Stem-loop PEs can be tethered to Cas9 nickase to produce tethered PEs.
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Stem-loop PEs can be tethered to Cas9 nickase to produce tethered PEs.
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Stem-loop PEs can be tethered to Cas9 nickase to produce tethered PEs.
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Stem-loop PEs can be tethered to Cas9 nickase to produce tethered PEs.
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Stem-loop PEs can be tethered to Cas9 nickase to produce tethered PEs.
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Stem-loop PEs were generated by adding stem-loop aptamers at the 3-prime terminal of pegRNA.
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
Stem-loop PEs were generated by adding stem-loop aptamers at the 3-prime terminal of pegRNA.
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
Stem-loop PEs were generated by adding stem-loop aptamers at the 3-prime terminal of pegRNA.
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
Stem-loop PEs were generated by adding stem-loop aptamers at the 3-prime terminal of pegRNA.
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
Stem-loop PEs were generated by adding stem-loop aptamers at the 3-prime terminal of pegRNA.
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
Stem-loop PEs were generated by adding stem-loop aptamers at the 3-prime terminal of pegRNA.
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
Stem-loop PEs were generated by adding stem-loop aptamers at the 3-prime terminal of pegRNA.
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Stem-loop PEs and tethered PEs increased small insertion, deletion, or point mutation editing efficiency by 2-fold to 4-fold on average in HEK293, U2OS, and HeLa cells.
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
editing efficiency fold increase 2-4 fold
Stem-loop PEs and tethered PEs increased small insertion, deletion, or point mutation editing efficiency by 2-fold to 4-fold on average in HEK293, U2OS, and HeLa cells.
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
editing efficiency fold increase 2-4 fold
Stem-loop PEs and tethered PEs increased small insertion, deletion, or point mutation editing efficiency by 2-fold to 4-fold on average in HEK293, U2OS, and HeLa cells.
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
editing efficiency fold increase 2-4 fold
Stem-loop PEs and tethered PEs increased small insertion, deletion, or point mutation editing efficiency by 2-fold to 4-fold on average in HEK293, U2OS, and HeLa cells.
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
editing efficiency fold increase 2-4 fold
Stem-loop PEs and tethered PEs increased small insertion, deletion, or point mutation editing efficiency by 2-fold to 4-fold on average in HEK293, U2OS, and HeLa cells.
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
editing efficiency fold increase 2-4 fold
Stem-loop PEs and tethered PEs increased small insertion, deletion, or point mutation editing efficiency by 2-fold to 4-fold on average in HEK293, U2OS, and HeLa cells.
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
editing efficiency fold increase 2-4 fold
Stem-loop PEs and tethered PEs increased small insertion, deletion, or point mutation editing efficiency by 2-fold to 4-fold on average in HEK293, U2OS, and HeLa cells.
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
editing efficiency fold increase 2-4 fold
Approval Evidence
1 source3 linked approval claimsfirst-pass slug split-pegrna-prime-editors
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility
Source:
design rationalesupports
The 3-prime extension of pegRNAs could negatively affect pegRNA stability or folding and compromise prime editing activity.
The 3’-extension of pegRNAs could negatively affect its stability or folding and comprise the PE activity.
Source:
engineering strategysupports
The modified pegRNAs were split into sgRNA and prime RNA to create split pegRNA prime editors.
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
Source:
functional propertysupports
Split pegRNA prime editors maintain prime editing activity and increase flexibility.
The resulting split pegRNA prime editors (SnPEs) maintain the PE activity and increase flexibility.
Source:
Comparisons
Source-backed strengths
The cited study reports that SnPEs maintain prime editing activity despite splitting the modified pegRNA. A further reported advantage is increased design flexibility relative to the unsplit pegRNA format.
Source:
which can be tethered to Cas9 nickase resulting in tethered PEs (tPEs)
Source:
Here we generated stem-loop PEs (sPEs) by adding stem-loop aptamers at the 3’-terminal of pegRNA
Source:
We split the modified pegRNAs into sgRNA and prime RNA. The resulting split pegRNA prime editors (SnPEs)
Source:
sPEs and tPEs increased the small insertion, deletion or point mutations efficiency by 2-4-fold on average in HEK293, U2OS and HeLa cells.
Compared with intron-containing CRISPRa construct
split pegRNA prime editors and intron-containing CRISPRa construct address a similar problem space because they share editing, recombination.
Shared frame: same top-level item type; shared target processes: editing, recombination
Compared with microfluidic organ-on-chip platforms
split pegRNA prime editors and microfluidic organ-on-chip platforms address a similar problem space because they share editing, recombination.
Shared frame: same top-level item type; shared target processes: editing, recombination
Strengths here: looks easier to implement in practice.
Compared with tethered PEs
split pegRNA prime editors and tethered PEs address a similar problem space because they share editing, recombination.
Shared frame: same top-level item type; shared target processes: editing, recombination; shared mechanisms: prime editing
Ranked Citations
- 1.