Highlander Genome Project
The Short Tail Mutation and Variants
There are currently 5 known and identified variants that cause a short tail mutation in cats. But, with the increase in DNA testing, it has become evident that there are more short tail variants that have not yet been identified. These are referred to as being "Novel Variants". (Novel means New or unidentified)
A cat must be DNA tested for all 5 known short tail variants with NO copies of any variant in order to be referred to as having a "Novel Variant".
There are 4 variants that are referred to as "T-box" . These 4 variants were first discovered in the Manx Breed and are commonly referred to as the Manx gene or mutation.
The 4 known T-box variants are: c.998delT, c.1199delC, c.1169delC, and c.998_1014dup17delGCC .
Inheriting a T-box variant can result in 4 phenotypes: 1) Rumpy - No tail 2) Rumpy Riser -small amount of cartilage, fat pad, or vertebrae 3) Stumpy -longer than the Riser but shorter than a full length tail. 4) Full length tail
The 5th known short tail variant is the HES7. It is not a T-box variant and is often referred to as the Japanese Bobtail gene or mutation. The HES7 is not found in the Highlander.
The presence of a Novel Variant(s) has been proven in 3 bloodlines in the Highlander breed through DNA testing with No copies of the 5 known variants. There are Novel Variants suspected but not proven in other short tail breeds as well.
The First DNA Testing of The Highlander Breed For Short Tail Variants
In 2016, the TICA Highlander Breed Chair contacted Geneticist Dr Leslie Lyons of the University Of Missouri College of Veterinary Medicine and Surgery, to request DNA testing of 8 Highlander cats to determine which of the 5 short tail variants were present. At that time, the Lyons' Den lab at MU was the only lab with the ability to test for short tail variants. Dr Lyons agreed to do the buccal swab tests at a cost of only $60 per cat (total $480). The results of the DNA tests showed that 5 of the Highlanders had 1 copy of the c.998delT variant and 3 Highlanders had 1 copy of the c.1199delC variant. For several years, it was assumed that the majority of Highlander cats were heterozygous for one of these T-box variants. T-box variants are "heterozygous dominant " which means a short tail cat can have 1 copy of a T-box variant and 1 copy of the "wild expression" (long tail) variant and will most likely have a shortened tail. Kittens inherit 2 tail variants, 1 from each parent. It is presumed that cats cannot be "homozygous" for the T-box variants which means they cannot have 2 copies. Therefore, cats with a T-box variant can still produce kittens with long tails when both parents pass along the variant for the "wild expression" (long tail) and not the T-box variant.
A standard Highlander is born with a naturally short tail. Long tail Highlanders are referred to as non-standard.
Researching Novel Variants
In Aug of 2020, I was very fortunate to adopt my first short tail Highlander with a "possible" Novel Variant. I named her Tomi LaRin of Templepantera. Her Sire, Inkadinka of Crabcreekcats, has a natural short tail and had been DNA tested for 4 of the 5 known short tail variants with NO copies found. I also had Tomi LaRin DNA tested by O.S. Wisdom Panel. She, also, has No copies of the common variants. *(PLEASE NOTE: N/N is the notation which indicates that No copies of the variants were found and also suggests that the cat is LIKELY to have the "wild expression" which is a LONG TAIL. The notation "N/N" is NOT a tail variant. Cats born with a shortened tail with N/N test results for the common variants a referred to as having A Possible Novel Variant)*
Tomi LaRin's mother is a long tail Highlander, Crabcreekcats Nyla Fidelia. Therefore, Tomi and her brother Binx could only have inherited their Sire's Possible Novel Variant. My first step was to DNA test for the 5th known short tail variant (c.998_1014dup17delGCC ). This test was completed on Tomi LaRin in Dec 2020 along with 2 Highlanders from Wildnwonderful Highlanders. The results were that No copies of the final known variant were found in any of the 3 Highlanders!
These 3 Highlander bloodlines have DNA PROOF of having a Novel Variant(s). Since the Novel Variant(s) have not yet been identified, it is not known if the 3 cats have the same Novel Variant(s).
The Highlander Genome Project Begins
From Aug through Nov 2020, I contacted several Labs requesting a DNA test for the c.998_1014dup17delGCC . I was referred to Dr Leslie Lyons by every laboratory. With the help of my well-known veterinarian and breeding specialist, Dr Marty Greer, I was able to get the attention of Dr Lyons who is the foremost authority on feline genetics. She is the head of the Lyon's Den Research Laboratory at the University Of Missouri College Of Veterinary Medicine and Surgery. Dr Lyons agreed to DNA test a group of Highlanders for the short tail variant c.998_1014dup17delGCC so long as the cats had already been DNA tested with a Possible Novel Variant. The cost for the test was $100 per cat. Dr Lyons was adamant that the DNA testing would be offered ONCE only as her lab IS NOT a commercial lab. I shared this information and the emails from Dr Lyons with the other Highlander breeders that had cats with a Possible Novel Variant. I answered questions and began coordinating the buccal swab testing for whomever wanted to DNA test their cats for the final variant. I informed everyone that they would need to submit buccal swabs to me with their cat's information as well as a $100 donation to the research lab per cat. Dr Lyons required all the samples to be sent together and all cat's information put on 1 submission form. She would then provide 1 written report which would include the test results for each cat. Samantha of Wildnwonderful Highlanders in WV volunteered to test 2 of her cats from different bloodlines. Along with my cat, we had just 3 cats volunteered. I submitted the samples from the 3 Highlanders on Dec 22, 2020. The cost of the DNA testing and shipping was at our own expense. Samantha and I split the total cost of $337.
We received the written DNA test results from Dr Lyons on Feb 4, 2021. All 3 Highlanders had 0 copies of the final variant! This is DNA Proof that these 3 Highlanders DO NOT have any of the 5 known short tail variants and officially have a Novel Variant(s). I shared this information with the other Highlander breeders who were interested in the project and posted the results in the TICA Highlander Breed Section Members Group. Also, the TICA board of directors requested updates on the project to be included in the Highlander Breed Section Reports submitted by our Breed Chair, Mary Hudec.
Along with the DNA report, Dr Lyons explained the next step in the process by which a new variant is identified. We would need to complete a Whole Genome Sequence (WGS) of the cats with a Novel Variant(s) at a cost of approximately $1000 per cat! Dr Lyons included the Highlanders in The 99 Lives Cat Initiative Genome Project under the heading HIGHLANDER TAIL STUDY . The 99 Lives Cat Initiative is a research project that Dr Lyons and her team are conducting in which they are Whole Genome Sequencing 99 cats of different breeds to identify variants responsible for feline diseases, traits, and mutations.
WHOLE GENOME SEQUENCING is used to create a DNA map of the millions of individual variants per cat. These DNA maps are examined in search of variant(s) that are strong suspects per disease, trait, or mutation. This is a long process that requires experienced geneticists sifting through MILLIONS of individual DNA variants! The 99 Lives Cat Initiative is a non-profit research project and is funded primarily by charitable donations.
In Feb 2021, Samantha of Wildnwonderful Highlanders agreed to be my assistant project coordinator for The Highlander Genome Project and the two of us began a series of fundraising campaigns. We needed to raise over $3000 in donations to fund the first trio of Highlanders to be WGS. All donations for the 99 Lives Cat Initiative Project are greatly appreciated. Tax deductible donations can be made online at: felinegenetics.missouri.edu Click on the 99 Lives Donate Now button and PLEASE type "Highlander Tail Study" in the box marked "Why are you donating" so the Highlanders receive credit for the fundraising. Thanks in advance for your support!
Blood Sample Trios
Searching for a Novel Variant through Whole Genome Sequencing requires a Trio of blood samples from the test subject (offspring) and both parents. The offspring and 1 of the parents must have the mutation while the other parent does not. The 3 bloodline Trios submitted for WGS are:
1) Crabcreekcats Tomi LaRin of TemplePantera Sire: Inkadinka of Crabcreekcats Dam: Crabcreekcats Nyla Fidelia
2) Wildnwonderful Wicca Sire: Wildnwonderful Fabio Dam: Davari of Wildnwonderful
3) NY City Slicker of Wildnwonderful Sire: Empirestatelynx Gunner Dam: Graceland of Empirestatelynx
In Feb 2021, Samantha and I had our vets collect the blood samples from our cats per Dr Lyons' instructions at our own expense. We had to set up the special shipping by FedEx Overnight Air at our own expense as well. The special shipping alone cost almost $100 per shipment, one from WV and one from WI plus the cost of the vet visits and supplies. Samantha also had a series of x-rays completed of the spines and tails of her cats to compare with x-rays of her cats with a known T-box variant at her own expense. Total cost $886. We contacted the owners of our cat's parents to request blood samples. In the meantime, Samantha and I continued our fundraising campaigns. My birthday fundraiser raised $550 in cash donations and another $140 in online donations made directly to The 99 Lives Cat Initiative on behalf of the Highlander Tail Study. Samantha raised $250 in online donations as part of a "Double Discount Donation" offered by her cattery where she gave double the amount of any donation as an adoption discount for her kittens with A Novel Tail Variant! We were also blessed with 2 online sponsorships of over $1000 each from my brothers, Bert Hultman and James Hultman.
I organized the collection and shipment of the blood samples from the parent cats by contacting their vets and using the cash donations I received to pay for the supplies, vet bills, and FedEx Overnight Air from New York, Texas, and Arkansas. Total cost of the parent cat's samples and shipping $599.
The Lyons Den Lab only sends out a bolus of blood samples for WGS about every 6 months. The blood samples from the parents of my cat, Tomi LaRin, arrived at the lab in time for the June 2021 WGS. But, we were still alittle short of our goal of $3000 in online donations to fund the WGS of this Trio. Once again we were blessed with support from wonderful Highlander breeders (2 of which did not even own a cat with A Novel Variant) Mary Hudec of Midwestern Highlanders, Audrey Bera of Bera Pantera Cattery, Shelly Foust of Stonehaven Highlanders, and Samantha Bradley of Wildnwonderful Highlanders. We surpassed our online goal for donations and Dr Lyons sent out the first Trio of blood samples for Whole Genome Sequencing.
After a few months, I checked in with the lab to ask if there was anything else they needed from us. Dr Lyons informed me that she would need buccal swabs from several cats related to the cats in the study. I contacted the Highlander breeders who had cats from the same blood lines as the 3 cats in the study. I also conducted a bake sale fundraiser to continue raising funds. In Nov 2021, I launched a T-shirt fundraiser with Custom Ink with 2 designs. "Highlander Cattitude" and "I Have Important Cat Stuff To Do". With help in spreading the word from Sue of Home Of The Highlander Cattery, Audrey of Bera Pantera Cattery, my sister in law, Emmy, our breed chair, Mary Hudec of Midwestern Highlanders, and other Highlander breeders and owners, the T-shirt fundraiser was a success! We raised $365.70.
Samantha, her sister Trista, and I collected more cash donations which I sent to 99 Lives Cat Initiative as an online donation on behalf of the Highlander Tail Study.
By Jan 2022, we had raised $4200 in online donations sent directly to The 99 Lives Cat Initiative on behalf of the Highlander Tail Study!!
I began organizing the collection and submission of buccal swab samples from several cats. As of Sept 2022, we have submitted buccal swab samples from 32 additional cats and kittens. Samples from 26 cats and kittens are related to the 3 bloodlines currently in The Highlander Tail Study. (Samples from related cats and kittens are essential to the study as Dr Lyons will be able to check for the "suspect variants" in the related cats to see if the suspect variant holds true. The DNA testing of these swabs are funded by the donations raised for the Highlander Genome Project.) The other 5 samples are from cats that are not related to the 3 bloodlines but have been DNA tested by O.S. Wisdom Panel with a Possible Novel Variant (these 5 cats have not been DNA tested for all 5 known variants and they will be DNA tested by Dr Lyons once A Novel Variant has been identified. Funding for these non-related cats has not yet been determined, but if we have raised enough in donations, these 5 will be funded by donations as well ). As Dr Lyons mentioned in her email regarding the submission of buccal swabs NOT RELATED to the bloodlines involved in the study, Quote: "DNA testing is not free". Please, keep in mind that we may have to raise more money for the samples that are being submitted that are NOT RELATED to the 3 bloodlines in the Highlander Tail Study. (Please consider sending a donation along with any buccal swabs from unrelated cats.)
We are hoping that several additional bloodlines have the same or similar Novel Variant(s). At this time we estimate approximately 8 different bloodlines from around the country and Canada.
Samantha of Wildnwonderful Highlanders, Trista of Bouwens Furbabies, and Shelly of Stonehaven Highlanders have several kittens that were born recently and are related to the 3 bloodlines in the study. Some of these kittens will be swabbed and submitted to the lab in the near future. Dr Lyons says "The More The Merrier" as related cats are important to verify that the suspected variant(s) hold true.
Please let me know if you are submitting buccal swabs from the 3 bloodlines. I am organizing the buccal swabs according to bloodlines and I am numbering the cats in order to keep the samples easily identifiable. Also, please, let me know if you plan to submit buccal swabs from any unrelated cats or kittens that have been tested for the common variants and have a Possible Novel Variant. If you are unsure if your cats are related, please ask their breeder for their bloodline. If you still do not know if your cats are related to the 3 bloodlines, then the cats must be listed as unrelated. Thank You for helping me help the lab by being organized. I also need to keep track of how much of the online donation funds we are using up.
Dr Lyons and her team at The Lyon's Den Lab are sifting through the MILLIONS of DNA variants in search of a candidate or "good suspect variant" that might be responsible for the short tail mutation. When they have a few good suspects they will compare DNA samples from the related cats to see if the suspect variant(s) hold true.
If she is satisfied with the results, then the unrelated cats can be tested as well. For now, we wait and let the experts do their thing. Samantha and I will continue fundraising for The 99 Lives Cat Initiative Project on behalf of the Highlanders.
Advice From The Experts and Breeders With Experience With A Variety Of Breeds
We are all VERY EXCITED about the discovery of A Novel Variant(s) in the Highlander breed and we are very hopeful that A Novel Variant(s) will be a completely new variant of a new mutation that can be homozygous. BUT we must be REALISTIC about this endeavor. It is not a good idea to make decisions based on what we HOPE the study will reveal. Please keep the current facts in mind when making choices for your breeding program.
1) We DO NOT know if any of the 3 bloodlines with proven Novel Variant(s) or any of the bloodlines with a Possible Novel Variant(s) are the same. At this time, we SHOULD NOT cross bloodlines with possibly different Novel Variants or breed a Novel Variant or Possible Novel Variant with any known short tail variant.
2) We DO NOT KNOW if the 3 Novel Variant(s) being studied by Dr Lyons or any of the Possible Novel Variants from other bloodlines are T-box, HES7, or a New mutation.
* If the study reveals that any of the Novel Variants are a new T-box variant, then the same information on the current T-box variants will apply to breeding with the new variant (nothing will have changed for a new T-box variant).
* If any of the Novel Variants are a NEW variant of the HES7, we might not be able to continue breeding with this new variant because the Japanese Bobtail Breed "Owns the rights" to the HES7 mutation.
3) We don't have enough documented information about any Novel Variant(s) to claim that any Novel Variant is safer or better than any other short tail variant. All of this information will be investigated once a Novel Variant(s) has been identified and studied.
Temple Pantera Cattery is committed to the Highlander Genome Project, the discovery of Novel Variant(s), the perpetuation of a homozygous mutation if discovered, and the advancement of the Highlander breed. BUT, we are NOT retiring our breeding cats with the c.998delT or the c.1199delC variants. The most up-to-date information on these variants along with our record of successful healthy litters and our cattery's success in the show ring suggests that our beautiful Highlanders are an asset to our breeding program and the future of Temple Pantera Cattery.
When I began my research, I discovered quite a bit of misinformation about short tail variants, including Novel Variant(s).
Here is a list of some CORRECT information. Please feel free to do your own research. I am always finding new information.
1) There could be MANY unidentified Novel Variants. Each bloodline could have a DIFFERENT Novel Variant(s) than the others.
2) The VGL lab at UC Davis is NOT researching Novel Short Tail Variant(s). This misinformation delayed my investigation of Novel Variant(s).
3) N/N is NOT a tail variant. N/N is the notation used by Optimal Selection to report NO copies of a variant. The Wisdom Panel by Optimal Selection does NOT test for all 5 known short tail variants. Cats with NO copies of ALL 5 known variants are referred to as having "A Novel Variant". Cats with N/N test results for only some of the known variants are referred to as having "A Possible Novel Variant".
4) There are 3 Highlanders that have been tested for all 5 known short tail variants with No copies of any of the 5 variants. Therefore, there are only 3 Highlander bloodlines with DNA proof of A Novel Variant . These cats are part of the Highlander Tail Study in the 99 Lives Cat Initiative Project being conducted by
Dr Leslie Lyons and her team at The Lyon's Den Lab located at MU.
The 3 bloodlines with DNA proof of A Novel Variant(s) are:
* Crabcreekcats Tomi LaRin of TemplePantera (and her Sire Inkadinka of Crabcreekcats)
* Wildnwonderful Wicca (and her Dam, Davari of Wildnwonderful)
* NY City Slicker of Wildnwonderful (and his Dam, Graceland of Empirestatelynx)
There are other Highlander bloodlines with a Possible Novel Variant. Buccal swab samples from unrelated bloodlines have been submitted to the Lyon's Den Lab and will be DNA tested once the lab has identified a Novel Variant in the bloodlines currently part of the study. As Dr Lyons mentioned in her email regarding the submission of buccal swabs not related to the bloodlines involved in the study, "DNA testing is not free". Please, keep in mind that we may have to raise more money for the samples that are being submitted that are NOT RELATED to the 3 bloodlines in the Highlander Tail Study. We are hoping that several additional bloodlines have the same or similar Novel Variant(s).
5) We CANNOT assume that all cats with A Novel Variant or a Possible Novel Variant have the SAME Variant. There may be several unidentified short tail variants yet to be discovered and identified.
6) We CANNOT assume that A Novel Variant is not a T-box variant! Novel Variant(s) could be any of 3 possible variant types:
a) a new variant of T-box b) a new variant of HES7 or c) a new variant of an unidentified short tail mutation.
7) The DNA tests conducted by Dr Lyons in 2016 were NOT a Genome Study. These were buccal swab DNA tests for the 5 short tail variants which Dr Lyons generously offered to provide at $60 per cat with a total cost of $480 for 8 cats. No cats Had A Novel Variant.
8) The length of the tail of a kitten with a T-box variant IS NOT determined by the length of the tails of it's parents. Therefore, breeding a long tail cat to a short tail cat DOES NOT increase the chance of medium length tails in the kittens. Also, breeding short tail to short tail DOES NOT increase the risk of producing rumpy kittens! Several studies have stated: "The length of the tail of the offspring is independent of the length of the tails of the parents. There are other factors, both biological and environmental, that influence the length of the tail of the offspring."
9) Abnormalities associated with Spina Bifida are found in both long tail and short tail breeds.
10) According to G.A.R.D. (the Genetic And Rare Diseases Info Center) Spina Bifida is a multifactorial condition with an undetermined etiology in humans and other species. Occurrences are sporadic and heredity has not been determined. This means that the exact cause or causes of Spina Bifida has not been identified. But, research has shown that Spina Bifida is a result of a COMBINATION of factors (biological, environmental, and nutritional) Therefore, there are NO tests to determine if an animal can "Carry" Spina Bifida. Some factors have been identified such as toxins and some chromosome abnormalities but many factors are still unknown. Please remember that Spina Bifida is multifactorial and the exact combination of factors is still being investigated. Based on this information, it is probable that short tail variants may not cause abnormalities associated with Spina Bifida without the presence of additional factors. As stated in the information I received from G.A.R.D. there must be multiple factors combined. Statistical Biologist Rachel Glover began The Manx Cat Genome Project on this premise. The goal of the study was to identify some of the "Other Factors" involved in determining the tail length and possibility of additional abnormalities.
Temple Pantera Cattery, as well as many other catteries, have been DNA testing, selectively breeding, investigating nutritional studies, providing proper nutrition, and adding nutritional supplements such as Folic Acid. Temple Pantera Cattery has never had an occurrence of abnormalities associated with Spina Bifida! Many other long tail and short tail breeds have also implemented these practices and have GREATLY reduced the occurrence of abnormalities associated with Spina Bifida within their breeds as well.
11) Some symptoms blamed on Spina Bifida and /or short tail variants are actually caused by other factors (such as parasites) and are unrelated to the presence of a short tail variant. Example: Urinary incontinence, fecal incontinence, and mega-colon can be caused by many other factors including Tritrichamonous Foetus, Giardia, Coxcidia, frequent UTIs, food allergies, toxins, other congenital defects, etc. Unfortunately, cats with a short tail are often misdiagnosed because of the assumption that the symptoms or problem is caused by the short tail variant.
12) The presence of kinks or curls in a cat's tail DOES NOT mean the cat has an abnormality associated with Spina Bifida. Several animal species, including some cat breeds, have shortened tails, kinks, and curls which are part of the breeds genetic make-up. A recent study confirmed that kinks and curls can be the result of a genetic variant in both long tail and short tail cats. Kinks and curls are required in the standards of the Japanese Bobtail and Kurulian Bobtail. Kinks and curls are allowed in the TICA Highlander standard, but not required.
13) There is no DNA evidence or proof of wild blood (such as the North American Bob Cat or Canadian Lynx) in the Highlander breed or it's predecessor/outcrossed breed known as the Highland Lynx. In fact, the newest version of the DNA test called the Wisdom Panel Complete includes tests for ancestry and has proven that there is NO WILD BLOOD in the Highlanders that have been tested for ancestry.
14) The name "Highlander" was adopted in 2005 as the breed was being developed separately from it's predecessor/outcrossed breed and the Highlander breed was recognized by TICA as an official pedigreed cat breed. The breed name was changed in order to identify the Highlander as a separate breed from it's predecessor/outcrossed breed and because the term "Lynx" was misleading as there is no proof of wild blood in either breed.