CRISPR

Benchmark
Report
An Inside Look at What’s
Happening at the Benchtop
TABLE OF CONTENTS
1. INTRODUCTION 3
2. OBJECTIVES 4
What Are CRISPR Researchers Working On?
Is CRISPR Hard?
Are CRISPR Researchers Satisfied With Their Results?

3. SURVEY RESULTS 6
CRISPR Researchers Have Professionally Diverse Profiles
Do-It-Yourself CRISPR Is Hard. Now We Know Why
CRISPR Researchers Are Not Satisfied With Their Results

4. CONCLUSION: DO-IT-YOURSELF CRISPR TAKES

ENORMOUS TIME AND COST INVESTMENT 23
5. APPENDIX 25
Survey Methodology
The Math: Time and Cost Calculations
 01 INTRODUCTION

INTRODUCTION

CRISPR genome editing is revolutionizing the way researchers can analyze, with unprecedented
precision and control, how genes are linked to biological function. CRISPR shows promise in
several areas, particularly in the field of personalized gene therapies.

We at Synthego are just as excited about the present and future applications of CRISPR as you
are, but we need to acknowledge that it is an emerging technology that is just starting to be
adopted within the workflows of academic and industry labs.

Making the best choice between a completely in-house CRISPR workflow or outsourcing all or
part of the effort can be confusing, what with all the noise in the marketplace. Plus, the workflow
is complex, with multiple factors impacting the outcome of CRISPR experiments. Any misstep
could cost months of wasted time and thousands of dollars.

We wanted to take a closer look at what CRISPR researchers are experiencing in their own labs
to see what we can learn from our scientific community. Accordingly, we surveyed 207 scientists
using CRISPR to find out about their challenges, applications, success levels, and satisfaction with
their experimental results. This report provides a detailed summary of the responses and patterns
that emerged from the survey.

We hope that this information will help researchers choose the best tools and realistically
estimate the time, labor, and cost investment required for a successful CRISPR journey.

3
 02 OBJECTIVES

OBJECTIVES

Most researchers take up the CRISPR challenge and embark on the do-it-yourself (DIY) route to
gain professional expertise in CRISPR. But CRISPR is complicated. Is the guide design right? How
many optimizations does my cell type need? Which transfection method should I use? These are
just a few of the questions that researchers need to grapple with when planning their CRISPR
experiment.

Yet, researchers have to balance complicated CRISPR workflows in the lab while managing their
project goals and deadlines.

We wanted to dig into this paradoxical

situation by getting an inside look at what’s A small misstep in the multi-step
workflow could result in weeks of
happening with CRISPR in today’s lab.
wasted time and money.
Therefore, we conducted a blind online
survey through various third party life
science publications. Broadly, our main
objectives behind the survey answered the
following three questions.

4
 02 OBJECTIVES

1. What Are CRISPR Researchers Working On?

What type of CRISPR edits are scientists making, and in what application and research areas?

The utility of CRISPR is expanding every day, and we wanted to understand the current trends in
the field. We investigated applications, researcher experience levels, research focus areas, and
editing methods to understand the profiles of CRISPR researchers.

2. Is CRISPR Hard?
What is the most challenging part of the CRISPR workflow? How much time are researchers
devoting to each step?

We wanted to learn about researchers’ experience of running a CRISPR experiment, so our survey
included questions regarding the details of the DIY CRISPR experimental workflow. We were
particularly interested in understanding how factors such as transfection method, optimization
levels, CRISPR formats, and cell types, to name a few, impact the outcome of the experiment.

3, Are CRISPR Researchers Satisfied With Their Results?

What average editing efficiencies are researchers achieving, and are they satisfied with their
results?

Besides understanding the challenges in a CRISPR experiment, we were also curious about the
perception of success among researchers. We asked if researchers are satisfied with the editing
efficiencies that they achieve in their CRISPR experiments. We also sought to determine the
ideal number of gene targets that they would like to study in parallel, as well their willingness to
outsource experiments while balancing their desire to master the CRISPR technique.

Our survey focused on the time and effort invested in experiments, but we also wanted to uncover
the true cost for researchers to execute the entire CRISPR workflow in their own lab. In the last
section in this report, we calculate the real cost of CRISPR based on data regarding hands-on
time, the duration for experimental completion, and the number of attempts before researchers
succeed.

For survey methodology, please see the Appendix.

5
 03 RESULTS - CRISPR RESEARCHERS ARE DIVERSE

CRISPR Researchers Have Professionally

Diverse Profiles

The survey respondents, in a nutshell, were professionally diverse. They ranged from graduate
students to executives, worked on a wide variety of applications and research areas, and
possessed CRISPR expertise, ranging from fewer than 6 months to more than 2 years. The
majority of the respondents worked in academia (66%), while 25% worked in industry settings,
such as biotechs and pharmaceuticals.

Wide Range in Roles

Respondents ranged from graduate student to executive level roles

The survey data showed that the

WhatisisYour
What YourCurrent
Current Role?
Role?
respondents belonged to diverse professional
roles, ranging from graduate students to Executive
Management
3.4%
director level roles. While a majority of the Research
Procurement
0.5%
Associate
respondents were principal investigators
Lab Principal
Investigator
(27.5%) and staff scientists (19.1%), the Manager
6.4%

graduate students’ category came in close at 7.4% 27.5%

16.7%. Director 7.8%

11.3%
Post-Doc
19.1%
16.7%
Staff
Scientist
Graduate
Student

6
 03 RESULTS - CRISPR RESEARCHERS ARE DIVERSE

Distributed Application Areas

Respondents reported working in a variety of application areas

What
What Application
Application AreaArea is Research
is Your Your Research
FocusedIn?
On?

23.2%

19.3%

13.0%

9.2%
8.2%
7.2% 6.8% 6.3%
4.8%
1.9%

Animal Disease Screening/ Primary Other Diagnostics Target iPS Pathway Safety/Tox
Models Models Target ID and Stem Validation Cells Analysis
Cells

We included general questions in our survey to get an understanding of the popular research
application areas.

CRISPR can be used to do genome editing in animal models at various developmental stages,
often to understand particular diseases. It came as no surprise that animal and disease modeling
applications, which bring research one step closer to clinical applications, together formed the
focus of 42.5% of the researchers.

The remaining respondents were spread across other application areas, such as screening and
target ID applications (13%) and primary and stem cell studies (9.2%). Recently, there has been a
surge in target validation (6.8%) applications, particularly in protein biology and immunotherapy
and in pathway analysis studies (4.8%) that elucidate the function and interplay of genes involved
in a biological pathway or specific disease.

7
 03 RESULTS - CRISPR RESEARCHERS ARE DIVERSE

Diverse Spectrum of Research Areas

Cancer and neuroscience among common research areas

What
What SpecificResearch
Specific Research Area
AreaAre
AreYou Studying?
You Studying?

18.0%

15.1%

12.2%

7.8%
6.8% 6.8% 6.3% 5.9%
4.4% 3.9% 3.4% 2.9% 2.4% 2.0% 2.0%

Cancer Neuroscience Basic Drug Immunology Cell and Dev Infectious Agriculture Therapeutics Other Immuno- Clinical Plant Metabolomics
Research Discovery Gene Biology Disease oncology Diagnostics Research
Therapy

The responses to the “Research Area” question were distributed across a wide spectrum of topics
indicating the diversity of areas in which CRISPR genome editing can play a role. Given the surge
of research in CAR T therapies and neurodegenerative disease research, it was no surprise that
cancer (18%) and neuroscience (15.1%) were the top two most studied research areas. Despite
CRISPR’s promising potential in cell and gene therapies and agriculture, basic research emerged
as a popular focus area among 12.2% of the survey participants.

Other interesting areas of focus included immunology, developmental biology, therapeutics, and
clinical diagnostics.

Resources
Curious about how CRISPR is being used in these application areas?
Check out our Application pages for descriptions of each of these areas.

CRISPR publications in different research areas are exploding.

Check out our Publications page to learn more.

8
 03 RESULTS - CRISPR RESEARCHERS ARE DIVERSE

Heterogeneity in Types of CRISPR Editing

Knockouts for the win; 50% of CRISPR researchers reported working on
knockouts

As CRISPR becomes a more common lab tool, its

What CRISPR Editing Method
What CRISPR Editing Method
Are You Using? utility is expanding. The addition of new methods
Are You Using?
to the CRISPR toolbox has enabled its expansion
Base DNA to base editing, DNA imaging, and diagnostics.
Editing Imaging
5.6% 2.6%
CRISPRi/
CRISPRa
But the classic knockout function that
7.7%
Detection/ made CRISPR popular still remains the
Diagnostics
8.2% method of choice for 50% of CRISPR
researchers.
Knock-in 12.8%
50.0%
(codon/SNV
change)
13.3% Knockout Knock-ins (single nucleotide variant (SNV)
changes and gene tags) comprised 26.1%,
Knock-in
(gene tag) while 7.7% of researchers reported working on
CRISPRi/CRISPRa experiments.

Defining the Methods

CRISPR Knockout Base Editing
A target gene is inactivated through a CRISPR-induced frameshift Method to switch out a single base in a DNA sequence, which is
or deletion. Ideal for gene function studies to determine if a gene particularly useful for developing treatments for single nucleotide
is linked to a specific cellular function or phenotype. polymorphisms (SNPs)

CRISPR Knock-in Detection/Diagnostics

Method to introduce an SNP or codon change. CRISPR knock-ins The specificity of CRISPR guide RNAs and nucleases can be
can also introduce gene tags for imaging studies, reporter assays, exploited for searching, rather than editing, thus showing
or downstream protein purification. promising potential in diagnostics.

CRISPRi/CRISPRa DNA Imaging

Method to silence or activate a target gene at the transcriptional A method in which modified Cas9 nuclease containing an imaging
level. CRISPRa can be used for gene overexpression studies, while probe (i.e., fluorescent protein) is introduced into cells along with
CRISPRi is used for gene repression experiments. a guide RNA to track genomic loci of interest in real time.

9
 03 RESULTS - CRISPR RESEARCHERS ARE DIVERSE

Cell Type Variety

Immortalized cell lines are the cell type of choice for 31.6% of researchers, but
primary and stem cells are catching up

What Cell Type Are You Working In?

Our survey data showed that immortalized

What Cell Type Are You Working In?
cell lines are a widely popular cell type
Whole Organism/
of choice with an average of 31.6% Embryo Immortalized
of researchers reportedly working in Cells

12.8%
immortalized cell lines.
31.6%

Stem 13.8%
These days, more and more researchers are Cells

now adopting stem cells and primary cells

as they are closer representatives of actual
18.8%
biological systems. The survey data backed
23.0%
up these trends, as stem cells and primary Other

cells together accounted for 36.8% of the Primary

Cells
respondents’ choices.

10
 03 RESULTS - CRISPR RESEARCHERS ARE DIVERSE

Importance of Mastering CRISPR

More than 60% of researchers, from both academia and industry, want to master
CRISPR as a professional skill

CRISPR is a revolutionary tool, and we know How Important is Mastering

How Important is Mastering CRISPR
that scientists want to hone their genome CRISPR To You?
To You?
editing technique by mastering this skill.
Accordingly, we asked respondents to rank
the importance of professionally mastering
67%
CRISPR. The data confirmed our hypothesis. 30%
3%

66% of respondents said it was

Very Somewhat Not Too
“very important” for them to master Important Important Important
CRISPR.

Interestingly, these stats were consistent even when we analyzed researchers in academia
(68.2%) and industry (62.7%) separately. These data reaffirm the importance of CRISPR skills
among all researchers in the field.

11
 03 RESULTS - DO-IT-YOURSELF CRISPR IS HARD

Do-It-Yourself CRISPR Is Hard

Now We Know Why

While CRISPR has promising potential and the scientific community is focused on its applications,
the issues that scientists face when dealing with CRISPR on a day-to-day basis are rarely given
as much attention. The major part of our survey investigated researchers’ experience with the
CRISPR workflow, including the time investment, success rate, and challenges in particular steps,
to name a few. As the results from the survey rolled in, one thing was clear—CRISPR is hard.

CRISPR is Tedious
Researchers spend an average of 61 hours of total hands-on time on a DIY
CRISPR experiment without accounting for the time spent on clones

The CRISPR workflow is tedious and labor-intensive. Planning and executing a successful CRISPR
experiment involves numerous steps, as each of them heavily influences the final outcome of the
experiment. On investigating the hands-on time that researchers spend on each step, the data
showed that researchers spend an average of 61 hours of hands-on time, without accounting
for the time spent on isolating clones.

HowMuch
How MuchHands-On
Hands-OnTime
Time is
is Required
Required Prior
Prior To
ToClonal
ClonalIsolation
Isolation Optimization
was the most
time-consuming
24 24 24 24 24

6.6 7 19.1 13.7 14.2

18

hours
6 18

hours
6 18

hours
6 18

hours
6 18

hours
6
step, accounting
12 12 12 12 12 for an average
Design Guide Optimization Transfection Analysis of 19.1 hours of
Preparation
researchers’ time.
31%
consider this step to be
the most challenging part
of the CRISPR workflow

Researchers also spent a significant amount of time on transfection (13.7 hours) and analysis (14.2
hours) steps.

12
 03 RESULTS - DO-IT-YOURSELF CRISPR IS HARD

The Real Time Invested in the CRISPR Workflow

Average experimental duration is 10 weeks, but researchers restart their
experiments an average of 7 times

While the average hands-on time for all the CRISPR steps is reportedly 61 hours, the process
itself is spread out over several weeks. Researchers report an average duration of 10 weeks to
complete the guide design, guide preparation, optimization, transfection, and analysis steps of the
CRISPR experiment. Screening and isolation of clones with the desired edit reportedly requires an
additional 9 weeks, on average. This is the best-case scenario if researchers succeed in their first
attempt.

Unfortunately, this does not become clear

Even a small misstep in any part of the
until researchers reach the final analysis stage
CRISPR workflow results in failed or
and can determine if the edit was successful.
delayed experiments.
In the case of sub-optimal editing efficiencies,

researchers are left with no choice but to repeat the whole experiment. Our survey results showed
that researchers restart their CRISPR experiments about 7 times, on average, before they obtain
their desired edit.

HowMany
How ManyTimes
Times
DoDo
YouYou Restart
Restart Your This means that the real-time investment
YourExperiment?
Experiment?
of CRISPR is 70 weeks, not 10 weeks, just
to obtain an edit.
29.2%

23.6%
The additional 9 weeks to take the successfully
edited cells to clone compounds the time
14.6%
13.5% 13.5%
investment even further. These numbers
indicate that a CRISPR experiment could easily
5.6%
cost researchers years of their time and effort,

0 1-2 3-4 5-7 8-10 >11

significantly impacting their publishing and/or
project course.

13
 03 RESULTS - DO-IT-YOURSELF CRISPR IS HARD

Choosing a Less-Than-Optimal Guide Format is Risky

Despite synthetic choices, more than 50% of researchers still rely on plasmids
and lentivirus

What Guide Format Are You Using?

Researchers can choose among the following
What Guide Format Are You Using?
formats of guide RNA for their experiment:
single guide RNA (sgRNA), plasmid, in vitro- IVT
4.1%
transcribed RNA, lentivirus, and cr:tracrRNA. Synthetic
cr:tracrRNA Plasmid
The choice of guide format is important as
it can largely influence the experimental 14.4% 34.9%

outcome.

Most of the researchers report using plasmids 18.5%

or lentivirus (53.4%) for their experiments.

Lentivirus
28.2%
While conventionally popular, these formats
are tedious, time-consuming, and prone to off-
Synthetic sgRNA
targets. Moreover, these methods often involve
the use of selection agents, such as antibiotics.

Synthetic sgRNA has recently gained

About 42% of CRISPR researchers
popularity as it is highly effective and results
who carry out clonal isolation use
in low off-target effects. No wonder more and
selection agents, although this
more researchers are switching to the sgRNA
method threatens the genomic
integrity of the cell. format. Among the CRISPR researchers in the
survey, 42.6% responded to using synthetic
guide RNA formats, either as sgRNA or two-
piece cr:tracrRNA.

Guide Formats 101

Plasmid: The DNA sequence encoding the desired guide RNA is incorporated in a plasmid
Synthetic sgRNA: Single guide RNA oligo of desired sequence is synthetically synthesized
Lentivirus: Similar to a plasmid system, guide RNA sequence is encoded in DNA form into lentivirus
cr:tracrRNA: Two-part system of the guide RNA, which is synthetically synthesized and can be easily ligated prior to use
IVT: In the in vitro-transcribed RNA format, guide RNA is enzymatically transcribed from the corresponding DNA sequence in a tube
14
 03 RESULTS - DO-IT-YOURSELF CRISPR IS HARD

The Transfection Disconnection

According to 31% of CRISPR users, transfection optimization is the most
challenging step

The next step to worry about is the

transfection step. Everyone knows that • 87% of CRISPR researchers conduct a
transfection optimization step.
guide design can only take you so far.
Optimizing transfection conditions for your
• They test an average of 7 transfection
particular cell type is also crucial to ensure
conditions with 4 gRNA sequences.
successful editing.

What
What StepDo
Step DoYou
You Find
Find the
theMost
MostChallenging?
Challenging?

Design 17.5%

Guide Preparation 5.5%

Optimization 30.8%

Transfection 9.9%

Analysis 8.8%

Clonal Isolation 27.5%

0% 20% 40%

Sometimes that’s enough, but sometimes it is not. Even after these optimizations, researchers
may end up with poor editing efficiencies. While Synthego offers a 200-point optimization to
ensure consistently high editing efficiencies for each cell type, performing a large number of
optimizations is challenging for labs that lack automation. Many researchers may thus not be able
to adequately assess their transfection parameters.

It comes as no surprise that 31% of CRISPR users consider transfection optimization to be

the most challenging component of the CRISPR workflow.

15
 03 RESULTS - DO-IT-YOURSELF CRISPR IS HARD

Analysis Paralysis
About 48% of researchers don’t analyze indels; those that do analyze them report
an average editing efficiency of 43%

How Do You Analyze Your Indels?

The primary quantitative measure of success
How Do You Analyze Your Indels?
in a CRISPR experiment is the average editing
Other efficiency (indel frequency). Web-based tools
Genomic
cleavage
assay 5.3% such as Inference of CRISPR Edits (ICE) may
7.4%
be used to analyze Sanger sequencing data
from edited cells to estimate the editing
Sanger-
16.8%
sequencing efficiency.
48.4%

22.1%
I do not
Not analyzing indels is a risky move.
analyze
indels
Researchers who skip this step rely on
Next-generation
sequencing non-genomic methods as validation.

A large proportion (48.4%) of researchers reported that they don’t even genotypically analyze their
edits, and move directly to subsequent assays.

Researchers who
WhatWhat
Is Your Average
Is Your AverageEditing Efficiency?
Editing Efficiency?
do measure their
100%

success report an
average editing
Industry: 48% efficiency of 43%,
Academic: 40%

indicating that
fewer than half the
target sequences

are edited. Surprisingly, the stats did not change much even when we examined just knockouts,
which are generally known to have better efficiencies than knock-ins. Moreover, the survey data
showed that editing efficiencies achieved by researchers in academia and industry settings
were comparable at 40% and 48%, respectively. Interestingly, editing efficiency stats were not
correlated with years of CRISPR expertise.

16
 03 RESULTS - DO-IT-YOURSELF CRISPR IS HARD

Challenges of Finding Clones With Desired Edits

Researchers screen an average of 137 clones

How
How ManyClones
Many Clones Do
DoYou
YouScreen?
Screen?

58.8%
Outsourcing to
Academic
53.3% Industry companies that
can offer efficiently
edited cells could
largely reduce the
25.3%
23.5% time spent on the
clonal isolation
10.7%
8.0% step.
5.9% 5.9% 5.9%
2.7%

0-100 101-200 201-300 301-400 401-500

Researchers noted that they screen about 137 cells on average in their experiment to identify
clones with the desired edit. While academic users screen 117 clones, researchers from industrial
settings reported screening an average of 206 cells to isolate clones with their desired edit.

These numbers are rather high, but quite expected if the editing efficiency is low. For comparison,
researchers who purchased a Synthego Knockout Cell Pool can screen just 20 clones on average
to obtain their desired edit, thanks to the high editing efficiencies (78%) of the pools.

Resources
Why is sgRNA increasingly being chosen over plasmids? Find out in this blog post.
Curious about the issues of using selectable markers in CRISPR? Learn more in this blog post.
Switching over from IVT to sgRNA format? Our Tips & Tricks guide has you covered!
Curious about our automated platform for optimizing CRISPR knockouts? Check out our
CRISPR Knockout Optimization application note!
Not sure which transfection method to choose? Here’s our online guide to help you decide.
Need reliable transfection protocols? Check out our Nucleofection, Electroporation, and Lipofection protocols.
Struggling with knock-ins? We’ve got you covered with our Tips & Tricks: Knock-ins resource.

17
 03 RESULTS - RESEARCHERS NOT SATISFIED WITH RESULTS

CRISPR Researchers Are Not Satisfied

With Their Results

While one objective of our survey was to understand the challenges that researchers face in
their DIY CRISPR workflows, we were also curious about their post-experiment status. Were
researchers satisfied with their editing efficiencies? Did they find the hard work worth it upon
crossing the finish line?

Long story short, the survey data says “no.” We elaborate on the data around researcher
satisfaction and speculate about the cause of their distress in the following section.

18
 03 RESULTS - RESEARCHERS NOT SATISFIED WITH RESULTS

Dealing With The Editing Efficiency Deficiency

Only 15% of researchers are “very satisfied” with their editing efficiencies

As noted in the previous section, researchers reported an average editing efficiency of 43%. The
number did not fluctuate drastically when comparing editing efficiencies of academic (40%) and
industry (48%) researchers.

How Satisfied Are You With Your Editing Efficiency?

How Satisfied Are You With Your Ediiting Efficiency? Given that researchers
48.4% spend several months,
on average, on their
29.5% CRISPR workflow, it is
14.7%
understandable that
6.3% only 14.7% of CRISPR DIY
1.1%
researchers said they were
“very satisfied” with their
Very Somewhat Neutral Somewhat Very editing efficiencies.
Dissatisfied Dissatisfied Satisfied Satisfied

Automation and engineering facilities enable Synthego to achieve 80% editing efficiencies in
different cell types, but these success levels are difficult to achieve consistently with limited
resources in the lab. It is no surprise that an even lower percentage of academic researchers
(12.2%) reported being “very satisfied” with their results. Researchers from industry seemed to fare
slightly better with 23.8% reporting being “very satisfied” with their average editing efficiencies.

It is worth noting from the data that an average of 48.4% of researchers, irrespective of whether
they are in academia or industry, are settling for editing efficiencies that they find only somewhat
satisfying.

19
 03 RESULTS - RESEARCHERS NOT SATISFIED WITH RESULTS

Settling For Fewer Gene Targets Than Desired

Researchers hope to study 17 gene targets in parallel, but settle for 7

How Many Gene Targets Do You Study?

How Many Gene Targets Do You Study?

Targeting multiple genes in

Ideal vs. Reality
parallel is not simple for labs
without automation. The
individual costs of all the
steps can quickly escalate
when targeting multiple gene
targets, making the process
Ideally, researchers In reality, researchers
would like to study only study on average
impractical.
17 targets 7 targets

CRISPR users reported that they would
ideally like to study 17 gene targets at a time.
Yet, the complications and time constraints
of the CRISPR workflow allow them to
investigate only 7 gene targets at a time.
This is not surprising, as targeting multiple
gene targets is not cost-free. While it might
seem prudent to test multiple genes in
the cell type optimization and transfection
steps, the hands-on time for other steps
adds up due to the individual complexities
and challenges each gene target presents.
The time and cost spent in guide RNA design,
genomic analysis, and clonal isolation steps, when
studying multiple gene targets, can compound to
a level where manual parallel processing becomes
difficult and impractical.
There is no single magic formula that works for
everyone. Every research team defines their
own “too much” and decides at what point they
might look to outsourcing of one or more steps
that allow them to study multiple gene targets in
parallel to expedite their CRISPR experiment.

20
 03 RESULTS - RESEARCHERS NOT SATISFIED WITH RESULTS

Walking The Tightrope

Researchers value skill development but struggle to balance it with time
pressures around projects

How Important is Mastering

CRISPR To You? As mentioned in a previous section, an
How Important Is Mastering CRISPR To You?
overwhelming majority (66%) of users
Academic Industry
reported that they consider it “very important”
3.3% 2.0% to master CRISPR skills professionally.

These numbers were consistent even when

28.5% 35.3% we dissected the data by institution type;
68.2% of academic researchers and 62.7% of
68.2% 62.7%
industry researchers considered CRISPR a very
Very Somewhat Not Too important skill to master, despite the time and
Important Important Important
cost it takes to perform a successful edit.

Are You Willing To Outsource On the other hand, researchers have to meet
AreYour
You CRISPR
Willing To Outsource Your
Experiment?
CRISPR Experiment? their project deadlines and/or accelerate their

Academic Industry
publications—a major career driver—while
learning on the go.

Given that CRISPR is hard and has a high

26.5% 31.6%
41.2% 36.8% failure rate, this becomes a challenging task.

32.4% 31.6% It’s no wonder that despite being strongly

motivated to learn CRISPR, 71% of researchers,
Yes Maybe No on average, are willing to outsource it.

21
 CRISPR Key Findings
Benchmark 2019 Edition
Report
An Inside Look at What’s Happening at the Benchtop

CRISPR Research is Diverse

Variety of Knockouts 2 out of 3

application are the most want to
areas common editing type master CRISPR

Do-It-Yourself CRISPR is Hard

31%
43%
7 attempts
to achieve a successful find optimization to be
Editing efficiency CRISPR-edited cell the most challenging step

Researchers Are Not Satisfied With Results

15%
report being
very satisfied studying 7 targets, however ideally
with their results would like to study 17 targets

The True Cost of CRISPR is Enormous

$15,340 + $3,054
spent on hands-on spent on reagents
labor cost

472 hours
resulting in
=
hands-on time
across all 7 attempts to $18,394
successfully complete
the CRISPR workflow total cost of a successful
CRISPR experiment
 04 CONCLUSION - DO-IT-YOURSELF CRISPR IS EXPENSIVE

Do-It-Yourself CRISPR Takes Enormous

Time and Cost Investment

Despite a majority of researchers finding it important to master the CRISPR technique, they are
not blind to its risks and associated time and cost investment. In the survey, a whopping 71% of
CRISPR DIYers said they would outsource their CRISPR experiments. When asked about the main
factor that would impact their decision, 46.1% noted their main concern was the price.

What Factors Are Are

What Factors Important
Important When Outsourcing
When Outsourcing Your Edit?
Your Edit?

Price 46.1%

Editing Efficiency 15.7%

Turnaround Time 14.6%

Edit Type 12.4%

Cell Type 11.2%

0% 20% 40%

This seems reasonable, but the “right
price” is rather subjective. In order to help
researchers put the true cost of DIY CRISPR
in perspective, we have calculated the cost of
a DIY CRISPR workflow.
Based on survey responses, we calculated
the typical costs, hands-on time, and
duration required to complete a successful
CRISPR editing experiment. We considered
the reported hands-on time that researchers
spend on each step of the CRISPR workflow,
duration of time required to complete the
experiment, the number of clones screened (if
any), and the number of failed attempts before
successfully generating their desired edit. Lab
reagent and personnel labor costs were assumed
based on academic and industry-standard labor
rates and typical vendor reagent costs. Capital
equipment and common wet laboratory costs
(e.g., incubators, pipette tips, cell culture plates)
were assumed in the cost and were not included
in the calculation. Refer to the Appendix for
complete cost analysis and assumptions.

23
 04 CONCLUSION - DO-IT-YOURSELF CRISPR IS EXPENSIVE

Based on these calculations, and assuming an average of six failed attempts,

we determined that it takes the average CRISPR DIYer:
• 472 hours of direct hands-on time to complete a successful CRISPR editing workflow
• 19 weeks to complete a successful experiment
• $15,340.00 in hands-on labor costs to generate a design, optimize, analyze, and isolate a
clone of the desired edited cell
• $891.31 in standard reagent costs, amounting to $18,394.19 in total costs to complete a
successful experiment
• 7 experimental attempts before generating the desired edit
(Note: calculations above factor in these repeated attempts)

What is the True Cost of Do-It-Yourself CRISPR?

Average

1 attempt 4 attempts 7 attempts 10 attempts

Time To Pool (Hours) 61 244 427 610

Cost To Pool $2,342.98 $9,371.92 $16,400.86 $23,429.80

Time To Clone (Hours) 45 45 45 45

Cost To Clone $1,993.33 $1,993.33 $1,993.33 $1,993.33

Total Time 106 289 472 655

Total Cost $4336.31 $11365.25 $18,394.19 $25,423.13

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 05 APPENDIX

Appendix I: Survey Methodology

We conducted a blind online survey through various third-party life science publications that cater
to a scientific audience. The survey contained 36 questions, and respondents who did not have
any CRISPR experience were automatically disqualified. Survey respondents received a $25 gift
card for their participation.

For analyses, we received 207 responses to the survey from researchers that have either
previously done or are currently doing CRISPR experiments.

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 05 APPENDIX

Appendix II: The Math:

Time and Cost Calculations

REAGENT COSTS
# rxns Cost per
Step Reagent Cost Unit size Amount/rxn Cost/rxn required experiment TOTAL REAGENT COST
Transfection CRISPRMax $180.00 100 2 $3.60 13 $46.80
Culture/Cloning Media (DMEM + FBS) $70.00 1 1 $70.00 2 $140.00
Edit Transfection Plasmid + sgRNA + Cas9 $0.00 0 0 $0.00 0 $0.00
Analysis DNA extraction $168.00 50 1 $3.36 13 $43.68
Analysis Sanger sequencing $5.00 1 1 $5.00 26 $130.00 $360.48 Pool
Cloning Media (DMEM + FBS) $70.00 1 1 $70.00 2 $140.00
Analysis DNA extraction $355.00 96 1 $3.70 40 $148.00
Clonal Isolation
Analysis Clean Up Kit $103.00 96 1 $1.07 40 $42.80
Analysis Sanger sequencing $5.00 1 1 $5.00 40 $200.00 $530.80 Clone
$891.28 Total clone

LABOR COSTS
# of Hours Cost (Hours x $/hour)*
Desgin 7 $227.50
Synthesis 7 $227.50
Optimization 19 $617.50
Tranfection 14 $455.00 Time (pool) Labor (pool)
Analysis 14 $455.00 61 $1,982.50
Clone 45 $1,462.50 Time (clone) Labor (clone)
106 $3,445.00

COST PER ATTEMPT

Time x Total TOTAL
# attempts Time (hours) attempts Hourly rate* Labor Cost Reagents Reagents COST
Pool 1 61 61 $32.50 $1,982.50 $360.48 $360.48 $2,342.98 Cost of POOL / attempt
Clone 1 45 45 $32.50 $1,462.50 $530.80 $530.80 $1,993.30
Total 1 106 106 $32.50 $3,445.00 $891.28 $891.28 $4,336.28 Cost of CLONE / attempt

COST PER SUCCESSFUL EXPERIMENT (6 attempts + 1 successful)*

Time x Total TOTAL
# attempts Time (hours) attempts Hourly rate* Labor Cost Reagents Reagents COST
Pool 7 61 427 $32.50 $13,877.50 $360.48 $2,523.36 $16,400.86 Cost of POOL / expt
Clone 1 45 45 $32.50 $1,462.50 $530.80 $530.80 $1,993.30
Total 8 106 472 $65.00 $15,340.00 $891.28 $3,054.16 $18,394.16 Cost of CLONE / expt

*ASSUMPTIONS FOR CALCULATIONS

Hourly wage for labor is $32.50
Assumptions for Calculations
Number of attempts based on results obtained from The Benchtop Benchmark Report: 2019 edition

• Hourly wage for labor is $32.50

• Number of attempts based on results obtained from The CRISPR Benchmark Report: 2019 Edition

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